LOCUS       Exported                8123 bp ds-DNA     circular SYN 05-FEB-2021
DEFINITION  pFuTraP vector expressing human ASCT2 with d7 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..8123
                     /organism="recombinant plasmid"
                     /lab_host="Gallus gallus"
                     /mol_type="other DNA"
     CDS             1..1608
                     /codon_start=1
                     /label=hASCT2-d7
                     /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV
                     RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII
                     LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN
                     ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYSTTYEEGGVKVPVGQEVE
                     GMNILGLVVFAIVFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFLVA
                     GKIVEMEDVGLLFARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTPLA
                     TAFGTSSSSATLPLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIAQL
                     SQQSLDFVKIITILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDWLV
                     DRSCTVLNVEGDALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPLLK
                     HYRGPAGDATVASEKESVM"
     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    631..636
                     /label=d7
     misc_feature    679..729
                     /label=TM4c
                     /note="Garaeva et al., 2018"
     misc_feature    745..861
                     /label=TM5
                     /note="Garaeva et al., 2018"
     misc_feature    874..972
                     /label=TM6
                     /note="Garaeva et al., 2018"
     misc_feature    982..1035
                     /label=HP1a
                     /note="Garaeva et al., 2018"
     misc_feature    1042..1083
                     /label=HP1b
                     /note="Garaeva et al., 2018"
     misc_feature    1099..1140
                     /label=TM7a
                     /note="Garaeva et al., 2018"
     misc_feature    1147..1194
                     /label=TM7b
                     /note="Garaeva et al., 2018"
     misc_feature    1216..1260
                     /label=HP2a
                     /note="Garaeva et al., 2018"
     misc_feature    1285..1323
                     /label=HP2b
                     /note="Garaeva et al., 2018"
     misc_feature    1348..1467
                     /label=TM8
                     /note="Garaeva et al., 2018"
     CDS             1624..1638
                     /codon_start=1
                     /label=GGGGS linker
                     /translation="GGGGS"
     CDS             1639..2355
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     misc_feature    2373..2959
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2960..3910
                     /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             4024..4409
                     /label=dLTR
     V_segment       complement(4410..7153)
                     /label=pLitmus 28i
     promoter        complement(4465..4483)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4504..4520)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4528..4544
                     /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(4552..4582)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4597..4618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4679..5267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5378..5887
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5929..6789)
                     /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(6790..6894)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7115..7131
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7135..7153
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             7154..7545
     misc_feature    7546..7563
                     /label=PBS
                     /note="the same as for RCAS"
     misc_feature    7600..7704
                     /label=packaging signal
                     /note="according to Banks JD and Linial ML, J. Virol., 
                     2000"
     gene            7816..8078
                     /label=p19
                     /note="p19"
     gene            8079..8092
                     /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 actctaccac ctatgaagag ggaggagtta aggtgcccgt ggggcaggag
      661 gtggagggga tgaacatcct gggcttggta gtgtttgcaa ttgtctttgg tgtggcgctg
      721 cggaagctgg ggcctgaagg ggagctgctt atccgcttct tcaactcctt caatgaggcc
      781 accatggttc tggtctcctg gatcatgtgg tacgcccctg tgggcatcat gttcctggtg
      841 gctggcaaga tcgtggagat ggaggatgtg ggtttactct ttgcccgcct tggcaagtac
      901 attctgtgct gcctgctggg tcacgccatc catgggctcc tggtactgcc cctcatctac
      961 ttcctcttca cccgcaaaaa cccctaccgc ttcctgtggg gcatcgtgac gccgctggcc
     1021 actgcctttg ggacctcttc cagttccgcc acgctgccgc tgatgatgaa gtgcgtggag
     1081 gagaataatg gcgtggccaa gcacatcagc cgtttcatcc tgcccatcgg cgccaccgtc
     1141 aacatggacg gtgccgcgct cttccagtgc gtggccgcag tgttcattgc acagctcagc
     1201 cagcagtcct tggacttcgt aaagatcatc accatcctgg tcacggccac agcgtccagc
     1261 gtgggggcag cgggcatccc tgctggaggt gtcctcactc tggccatcat cctcgaagca
     1321 gtcaacctcc cggtcgacca tatctccttg atcctggctg tggactggct agtcgaccgg
     1381 tcctgtaccg tcctcaatgt agaaggtgac gctctggggg caggactcct ccaaaattac
     1441 gtggaccgta cggagtcgag aagcacagag cctgagttga tacaagtgaa gagtgagctg
     1501 cccctggatc cgctgccagt ccccactgag gaaggaaacc ccctcctcaa acactatcgg
     1561 gggcccgcag gggatgccac ggtcgcctct gagaaggaat cagtcatgac tagtaccggt
     1621 accggaggtg gtggatccgt gagcaagggc gccgagctgt tcaccggcat cgtgcccatc
     1681 ctgatcgagc tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag
     1741 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgcct
     1801 gtgccctggc ccaccctggt gaccaccctg agctacggcg tgcagtgctt ctcacgctac
     1861 cccgatcaca tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacatccag
     1921 gagcgcacca tcttcttcga ggatgacggc aactacaagt cgcgcgccga ggtgaagttc
     1981 gagggcgata ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc
     2041 aacatcctgg gcaataagat ggagtacaac tacaacgccc acaatgtgta catcatgacc
     2101 gacaaggcca agaatggcat caaggtgaac ttcaagatcc gccacaacat cgaggatggc
     2161 agcgtgcagc tggccgacca ctaccagcag aataccccca tcggcgatgg ccctgtgctg
     2221 ctgcccgata accactacct gtccacccag agcgccctgt ccaaggaccc caacgagaag
     2281 cgcgatcaca tgatctactt cggcttcgtg accgccgccg ccatcaccca cggcatggat
     2341 gagctgtaca agtgagcggc cgcgactcta gacccctctc cctccccccc ccctaacgtt
     2401 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     2461 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     2521 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     2581 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     2641 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     2701 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     2761 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     2821 cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt ttagtcgagg
     2881 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     2941 tgataatatg gccacaacca tggcggaagg atccgtcgcc aggcagcctg acctcttgac
     3001 ctgcgacgat gagccgatcc atatccccgg tgccatccaa ccgcatggac tgctgctcgc
     3061 cctcgccgcc gacatgacga tcgttgccgg cagcgacaac cttcccgaac tcaccggact
     3121 ggcgatcggc gccctgatcg gccgctctgc ggccgatgtc ttcgactcgg agacgcacaa
     3181 ccgtctgacg atcgccttgg ccgagcccgg ggcggccgtc ggagcaccga tcactgtcgg
     3241 cttcacgatg cgaaaggacg caggcttcat cggctcctgg catcgccatg atcagctcat
     3301 cttcctcgag ctcgagcctc cccagcggga cgtcgccgag ccgcaggcgt tcttccgccg
     3361 caccaacagc gccatccgcc gcctgcaggc cgccgaaacc ttggaaagcg cctgcgccgc
     3421 cgcggcgcaa gaggtgcgga agattaccgg cttcgatcgg gtgatgatct atcgcttcgc
     3481 ctccgacttc agcggcgaag tgatcgcaga ggatcggtgc gccgaggtcg agtcaaaact
     3541 aggcctgcac tatcctgcct caaccgtgcc ggcgcaggcc cgtcggctct ataccatcaa
     3601 cccggtacgg atcattcccg atatcaatta tcggccggtg ccggtcaccc cagacctcaa
     3661 tccggtcacc gggcggccga ttgatcttag cttcgccatc ctgcgcagcg tctcgcccgt
     3721 ccatctggag ttcatgcgca acataggcat gcacggcacg atgtcgatct cgattttgcg
     3781 cggcgagcga ctgtggggat tgatcgtttg ccatcaccga acgccgtact acgtcgatct
     3841 cgatggccgc caagcctgcg agctagtcgc ccaggttctg gcctggcaga tcggcgtgat
     3901 ggaagagtga cgcgtatctg aggggactag ggtgtgttta ggcgaaaagc ggggcttcgg
     3961 ttgtacgcgg ttaggagtcc cctcgaggta tggcagatat gcttttgcat agggaggggg
     4021 aaatgtagtc ttaatcatag gttaacatgt atattaccaa ataagggaat cgcctgatgc
     4081 accaaataag gtattatatg atcccattgg tggcgaggga gcgacctgag ggcatatggg
     4141 cgttaacaga actgtctgtc cttgcgtcat tccttcctca tcggatcatg tacgcggcag
     4201 agtatgattg gataacagga tggcaccatt catcgtggcg catgctgatt ggtgcgacta
     4261 aggagttatg taacccacga atgtacttaa gcttgtagtt gctaacaata aagggccatt
     4321 ctacctctca ccacattggt gtgcacctgg gttgatggcc ggaccgtcga ttccctgacg
     4381 actacgagca cctgaatgaa gcagaaggca tcctgcagga attcctcgag acgtacggtg
     4441 cgcgcgatgc attcgaagat ctgccctata gtgagtcgta ttacgtagct tggcgtaatc
     4501 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4561 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaca
     4621 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4681 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4741 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4801 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4861 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4921 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4981 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5041 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5101 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5161 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5221 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5281 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5341 tgagattatc aaaaaggatc ttcacctaga tccttttacg cgccctgtag cggcgcatta
     5401 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5461 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgctttccc cgtcaagctc
     5521 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5581 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5641 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5701 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5761 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     5821 ttacaattta aatatttgct tatacaatct tcctgttttt ggggcttttc tgattatcaa
     5881 ccggggtaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5941 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6001 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6061 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6121 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6181 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6241 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6301 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6361 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6421 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6481 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6541 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaga
     6601 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6661 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6721 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6781 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6841 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6901 tccccgaaaa gtgccacctg acgtagttaa caaaaaaaag cccgccgaag cgggctttat
     6961 taccaagcga agcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7021 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     7081 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tccgtaatac
     7141 gactcactat aggtgtagtc ttaatcatag gttaacatgt atattaccaa ataagggaat
     7201 cgcctgatgc accaaataag gtattatatg atcccattgg tggcgaggga gcgacctgag
     7261 ggcatatggg cgttaacaga actgtctgtc cttgcgtcat tccttcctca tcggatcatg
     7321 tacgcggcag agtatgattg gataacagga tggcaccatt catcgtggcg catgctgatt
     7381 ggtgcgacta aggagttatg taacccacga atgtacttaa gcttgtagtt gctaacaata
     7441 aagggccatt ctacctctca ccacattggt gtgcacctgg gttgatggcc ggaccgtcga
     7501 ttccctgacg actacgagca cctgaatgaa gcagaaggct tcatttggtg accccgacgt
     7561 gatcgttagg gaatagtggt cggccacagg cggcgtggcg atcctgtctc catccgtctc
     7621 gtcgaccggg aggcgagttc gatgaccctg atagaggggg ctgcggctta gggaggcaga
     7681 agctgagcac cgtcggtagg agctccaggg cccggagcga ctgacccctg ccgagaactc
     7741 agagggtcgt cggaagacgg agagtgagcc cgacgaccac cccaggcacg tctttggtcg
     7801 gcctgcggat caagctagca agccgtcatt aaggtgattt cgtccgcgtg taaaacctat
     7861 tgcgggaaaa tctctccttc taagaaggaa ataggggcca tgttgtccct gttacaaaag
     7921 gaagggttgc tttagtctcc ctcagattta tattctccgg ggtcctggga tcccatcact
     7981 gcggcgctct cccagcgggc aatggtactt ggaaaatcgg gagagttaaa aacctgggga
     8041 ttggttttgg gggcattgaa ggcggctcga gagtaacagg actaacttga atcgatgcgc
     8101 taccggactc agatcttgcc acc
//
