LOCUS       Exported                8093 bp ds-DNA     circular SYN 05-FEB-2021
DEFINITION  pFuTraP vector expressing human ASCT2 with d17 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8138)
  AUTHORS   Stafl K, Travnicek M, Kucerova D, Pecnova L, Krchlikova V, Galikova 
            E, Hejnar J and Trejbalova K
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 5, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8093
                     /organism="recombinant plasmid"
                     /lab_host="Gallus gallus"
                     /mol_type="other DNA"
     CDS             1..1578
                     /codon_start=1
                     /label=hASCT2-d17
                     /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV
                     RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII
                     LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN
                     ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYSGGGQEVEGMNILGLVVF
                     AIVFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFLVAGKIVEMEDVG
                     LLFARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTPLATAFGTSSSSA
                     TLPLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIAQLSQQSLDFVKI
                     ITILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDWLVDRSCTVLNVE
                     GDALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPLLKHYRGPAGDAT
                     VASEKESVM"
     misc_feature    124..150
                     /label=TM1a
                     /note="Garaeva et al., 2018"
     misc_feature    154..222
                     /product="Garaeva et al., 2018"
                     /label=TM1b
     misc_feature    247..351
                     /label=TM2
                     /note="Garaeva et al., 2018"
     misc_feature    376..459
                     /label=TM3
                     /note="Garaeva et al., 2018"
     misc_feature    460..540
                     /label=region N
     misc_feature    541..570
                     /product="Garaeva et al., 2018"
                     /label=TM4a
     misc_feature    583..609
                     /label=TM4b
     misc_feature    616..621
                     /label=d17
     misc_feature    649..699
                     /label=TM4c
                     /note="Garaeva et al., 2018"
     misc_feature    715..831
                     /label=TM5
                     /note="Garaeva et al., 2018"
     misc_feature    844..942
                     /label=TM6
                     /note="Garaeva et al., 2018"
     misc_feature    952..1005
                     /label=HP1a
                     /note="Garaeva et al., 2018"
     misc_feature    1012..1053
                     /label=HP1b
                     /note="Garaeva et al., 2018"
     misc_feature    1069..1110
                     /label=TM7a
                     /note="Garaeva et al., 2018"
     misc_feature    1117..1164
                     /label=TM7b
                     /note="Garaeva et al., 2018"
     misc_feature    1186..1230
                     /label=HP2a
                     /note="Garaeva et al., 2018"
     misc_feature    1255..1293
                     /label=HP2b
                     /note="Garaeva et al., 2018"
     misc_feature    1318..1437
                     /label=TM8
                     /note="Garaeva et al., 2018"
     CDS             1594..1608
                     /codon_start=1
                     /label=GGGGS linker
                     /translation="GGGGS"
     CDS             1609..2325
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     misc_feature    2343..2929
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2930..3880
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     LTR             3994..4379
                     /label=dLTR
     V_segment       complement(4380..7123)
                     /label=pLitmus 28i
     promoter        complement(4435..4453)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4435..4453)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4474..4490)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4498..4514
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4522..4552)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4567..4588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4649..5237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5348..5857
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5899..6759)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6760..6864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7085..7101
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7105..7123
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7105..7123
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             7124..7515
     LTR             7124..7509
                     /label=dLTR
     misc_feature    7516..7533
                     /label=PBS
                     /note="the same as for RCAS"
     misc_feature    7570..7674
                     /label=packaging signal
                     /note="according to Banks JD and Linial ML, J. Virol., 
                     2000"
     gene            7786..8048
                     /label=p19
                     /note="p19"
     gene            8049..8062
                     /label=fragment p27
                     /note="fragment p27"
ORIGIN
        1 atggtggccg atcctcctcg agactccaag gggctcgcag cggcggagcc caccgccaac
       61 gggggcctgg cgctggcctc catcgaggac caaggcgcgg cagcaggcgg ctactgcggt
      121 tcccgggacc aggtgcgccg ctgccttcga gccaacctgc ttgtgctgct gacagtggtg
      181 gccgtggtgg ccggcgtggc gctgggactg ggggtgtcgg gggccggggg tgcgctggcg
      241 ttgggcccgg agcgcttgag cgccttcgtc ttcccgggcg agctgctgct gcgtctgctg
      301 cggatgatca tcttgccgct ggtggtgtgc agcttgatcg gcggcgccgc cagcctggac
      361 cccggcgcgc tcggccgtct gggcgcctgg gcgctgctct ttttcctggt caccacgctg
      421 ctggcgtcgg cgctcggagt gggcttggcg ctggctctgc agccgggcgc cgcctccgcc
      481 gccatcaacg cctccgtggg agccgcgggc agtgccgaaa atgcccccag caaggaggtg
      541 ctcgattcgt tcctggatct tgcgagaaat atcttccctt ccaacctggt gtcagcagcc
      601 tttcgctcat actccggagg tgggcaggag gtggagggga tgaacatcct gggcttggta
      661 gtgtttgcaa ttgtctttgg tgtggcgctg cggaagctgg ggcctgaagg ggagctgctt
      721 atccgcttct tcaactcctt caatgaggcc accatggttc tggtctcctg gatcatgtgg
      781 tacgcccctg tgggcatcat gttcctggtg gctggcaaga tcgtggagat ggaggatgtg
      841 ggtttactct ttgcccgcct tggcaagtac attctgtgct gcctgctggg tcacgccatc
      901 catgggctcc tggtactgcc cctcatctac ttcctcttca cccgcaaaaa cccctaccgc
      961 ttcctgtggg gcatcgtgac gccgctggcc actgcctttg ggacctcttc cagttccgcc
     1021 acgctgccgc tgatgatgaa gtgcgtggag gagaataatg gcgtggccaa gcacatcagc
     1081 cgtttcatcc tgcccatcgg cgccaccgtc aacatggacg gtgccgcgct cttccagtgc
     1141 gtggccgcag tgttcattgc acagctcagc cagcagtcct tggacttcgt aaagatcatc
     1201 accatcctgg tcacggccac agcgtccagc gtgggggcag cgggcatccc tgctggaggt
     1261 gtcctcactc tggccatcat cctcgaagca gtcaacctcc cggtcgacca tatctccttg
     1321 atcctggctg tggactggct agtcgaccgg tcctgtaccg tcctcaatgt agaaggtgac
     1381 gctctggggg caggactcct ccaaaattac gtggaccgta cggagtcgag aagcacagag
     1441 cctgagttga tacaagtgaa gagtgagctg cccctggatc cgctgccagt ccccactgag
     1501 gaaggaaacc ccctcctcaa acactatcgg gggcccgcag gggatgccac ggtcgcctct
     1561 gagaaggaat cagtcatgac tagtaccggt accggaggtg gtggatccgt gagcaagggc
     1621 gccgagctgt tcaccggcat cgtgcccatc ctgatcgagc tgaatggcga tgtgaatggc
     1681 cacaagttca gcgtgagcgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1741 aagttcatct gcaccaccgg caagctgcct gtgccctggc ccaccctggt gaccaccctg
     1801 agctacggcg tgcagtgctt ctcacgctac cccgatcaca tgaagcagca cgacttcttc
     1861 aagagcgcca tgcctgaggg ctacatccag gagcgcacca tcttcttcga ggatgacggc
     1921 aactacaagt cgcgcgccga ggtgaagttc gagggcgata ccctggtgaa tcgcatcgag
     1981 ctgaccggca ccgatttcaa ggaggatggc aacatcctgg gcaataagat ggagtacaac
     2041 tacaacgccc acaatgtgta catcatgacc gacaaggcca agaatggcat caaggtgaac
     2101 ttcaagatcc gccacaacat cgaggatggc agcgtgcagc tggccgacca ctaccagcag
     2161 aataccccca tcggcgatgg ccctgtgctg ctgcccgata accactacct gtccacccag
     2221 agcgccctgt ccaaggaccc caacgagaag cgcgatcaca tgatctactt cggcttcgtg
     2281 accgccgccg ccatcaccca cggcatggat gagctgtaca agtgagcggc cgcgactcta
     2341 gacccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     2401 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     2461 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     2521 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     2581 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     2641 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     2701 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     2761 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     2821 tacacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     2881 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca tggcggaagg
     2941 atccgtcgcc aggcagcctg acctcttgac ctgcgacgat gagccgatcc atatccccgg
     3001 tgccatccaa ccgcatggac tgctgctcgc cctcgccgcc gacatgacga tcgttgccgg
     3061 cagcgacaac cttcccgaac tcaccggact ggcgatcggc gccctgatcg gccgctctgc
     3121 ggccgatgtc ttcgactcgg agacgcacaa ccgtctgacg atcgccttgg ccgagcccgg
     3181 ggcggccgtc ggagcaccga tcactgtcgg cttcacgatg cgaaaggacg caggcttcat
     3241 cggctcctgg catcgccatg atcagctcat cttcctcgag ctcgagcctc cccagcggga
     3301 cgtcgccgag ccgcaggcgt tcttccgccg caccaacagc gccatccgcc gcctgcaggc
     3361 cgccgaaacc ttggaaagcg cctgcgccgc cgcggcgcaa gaggtgcgga agattaccgg
     3421 cttcgatcgg gtgatgatct atcgcttcgc ctccgacttc agcggcgaag tgatcgcaga
     3481 ggatcggtgc gccgaggtcg agtcaaaact aggcctgcac tatcctgcct caaccgtgcc
     3541 ggcgcaggcc cgtcggctct ataccatcaa cccggtacgg atcattcccg atatcaatta
     3601 tcggccggtg ccggtcaccc cagacctcaa tccggtcacc gggcggccga ttgatcttag
     3661 cttcgccatc ctgcgcagcg tctcgcccgt ccatctggag ttcatgcgca acataggcat
     3721 gcacggcacg atgtcgatct cgattttgcg cggcgagcga ctgtggggat tgatcgtttg
     3781 ccatcaccga acgccgtact acgtcgatct cgatggccgc caagcctgcg agctagtcgc
     3841 ccaggttctg gcctggcaga tcggcgtgat ggaagagtga cgcgtatctg aggggactag
     3901 ggtgtgttta ggcgaaaagc ggggcttcgg ttgtacgcgg ttaggagtcc cctcgaggta
     3961 tggcagatat gcttttgcat agggaggggg aaatgtagtc ttaatcatag gttaacatgt
     4021 atattaccaa ataagggaat cgcctgatgc accaaataag gtattatatg atcccattgg
     4081 tggcgaggga gcgacctgag ggcatatggg cgttaacaga actgtctgtc cttgcgtcat
     4141 tccttcctca tcggatcatg tacgcggcag agtatgattg gataacagga tggcaccatt
     4201 catcgtggcg catgctgatt ggtgcgacta aggagttatg taacccacga atgtacttaa
     4261 gcttgtagtt gctaacaata aagggccatt ctacctctca ccacattggt gtgcacctgg
     4321 gttgatggcc ggaccgtcga ttccctgacg actacgagca cctgaatgaa gcagaaggca
     4381 tcctgcagga attcctcgag acgtacggtg cgcgcgatgc attcgaagat ctgccctata
     4441 gtgagtcgta ttacgtagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4501 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4561 tgcctaatga gtgagctaac tcacattaca tgtgagcaaa aggccagcaa aaggccagga
     4621 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4681 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4741 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4801 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4861 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4921 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4981 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5041 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5101 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5161 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5221 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5281 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5341 tccttttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5401 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5461 ctcgccacgt tcgctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5521 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     5581 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5641 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     5701 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5761 ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aatatttgct tatacaatct
     5821 tcctgttttt ggggcttttc tgattatcaa ccggggtaaa tcaatctaaa gtatatatga
     5881 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5941 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6001 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6061 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6121 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6181 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6241 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6301 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6361 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6421 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6481 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     6541 cagaacttta aaagtgctca tcattggaga acgttcttcg gggcgaaaac tctcaaggat
     6601 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6661 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6721 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6781 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6841 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtagttaa
     6901 caaaaaaaag cccgccgaag cgggctttat taccaagcga agcgccattc gccattcagg
     6961 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     7021 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     7081 cgttgtaaaa cgacggccag tccgtaatac gactcactat aggtgtagtc ttaatcatag
     7141 gttaacatgt atattaccaa ataagggaat cgcctgatgc accaaataag gtattatatg
     7201 atcccattgg tggcgaggga gcgacctgag ggcatatggg cgttaacaga actgtctgtc
     7261 cttgcgtcat tccttcctca tcggatcatg tacgcggcag agtatgattg gataacagga
     7321 tggcaccatt catcgtggcg catgctgatt ggtgcgacta aggagttatg taacccacga
     7381 atgtacttaa gcttgtagtt gctaacaata aagggccatt ctacctctca ccacattggt
     7441 gtgcacctgg gttgatggcc ggaccgtcga ttccctgacg actacgagca cctgaatgaa
     7501 gcagaaggct tcatttggtg accccgacgt gatcgttagg gaatagtggt cggccacagg
     7561 cggcgtggcg atcctgtctc catccgtctc gtcgaccggg aggcgagttc gatgaccctg
     7621 atagaggggg ctgcggctta gggaggcaga agctgagcac cgtcggtagg agctccaggg
     7681 cccggagcga ctgacccctg ccgagaactc agagggtcgt cggaagacgg agagtgagcc
     7741 cgacgaccac cccaggcacg tctttggtcg gcctgcggat caagctagca agccgtcatt
     7801 aaggtgattt cgtccgcgtg taaaacctat tgcgggaaaa tctctccttc taagaaggaa
     7861 ataggggcca tgttgtccct gttacaaaag gaagggttgc tttagtctcc ctcagattta
     7921 tattctccgg ggtcctggga tcccatcact gcggcgctct cccagcgggc aatggtactt
     7981 ggaaaatcgg gagagttaaa aacctgggga ttggttttgg gggcattgaa ggcggctcga
     8041 gagtaacagg actaacttga atcgatgcgc taccggactc agatcttgcc acc
//
