LOCUS       Exported                8105 bp ds-DNA     circular SYN 05-FEB-2021
DEFINITION  pFuTraP vector expressing human ASCT2 with d13 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..8105
                     /organism="recombinant plasmid"
                     /lab_host="Gallus gallus"
                     /mol_type="other DNA"
     CDS             1..1590
                     /codon_start=1
                     /label=hASCT2-d13
                     /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV
                     RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII
                     LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN
                     ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYSTTGGPVGQEVEGMNILG
                     LVVFAIVFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFLVAGKIVEM
                     EDVGLLFARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTPLATAFGTS
                     SSSATLPLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIAQLSQQSLD
                     FVKIITILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDWLVDRSCTV
                     LNVEGDALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPLLKHYRGPA
                     GDATVASEKESVM"
     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    622..627
                     /label=d13
     misc_feature    661..711
                     /label=TM4c
                     /note="Garaeva et al., 2018"
     misc_feature    727..843
                     /label=TM5
                     /note="Garaeva et al., 2018"
     misc_feature    856..954
                     /label=TM6
                     /note="Garaeva et al., 2018"
     misc_feature    964..1017
                     /label=HP1a
                     /note="Garaeva et al., 2018"
     misc_feature    1024..1065
                     /label=HP1b
                     /note="Garaeva et al., 2018"
     misc_feature    1081..1122
                     /label=TM7a
                     /note="Garaeva et al., 2018"
     misc_feature    1129..1176
                     /label=TM7b
                     /note="Garaeva et al., 2018"
     misc_feature    1198..1242
                     /label=HP2a
                     /note="Garaeva et al., 2018"
     misc_feature    1267..1305
                     /label=HP2b
                     /note="Garaeva et al., 2018"
     misc_feature    1330..1449
                     /label=TM8
                     /note="Garaeva et al., 2018"
     CDS             1606..1620
                     /codon_start=1
                     /label=GGGGS linker
                     /translation="GGGGS"
     CDS             1621..2337
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     misc_feature    2355..2941
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2942..3892
                     /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             4006..4391
                     /label=dLTR
     V_segment       complement(4392..7135)
                     /label=pLitmus 28i
     promoter        complement(4447..4465)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4486..4502)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4510..4526
                     /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(4534..4564)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4579..4600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4661..5249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5360..5869
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5911..6771)
                     /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(6772..6876)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7097..7113
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7117..7135
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             7136..7527
     misc_feature    7528..7545
                     /label=PBS
                     /note="the same as for RCAS"
     misc_feature    7582..7686
                     /label=packaging signal
                     /note="according to Banks JD and Linial ML, J. Virol., 
                     2000"
     gene            7798..8060
                     /label=p19
                     /note="p19"
     gene            8061..8074
                     /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 cggagggccc gtggggcagg aggtggaggg gatgaacatc
      661 ctgggcttgg tagtgtttgc aattgtcttt ggtgtggcgc tgcggaagct ggggcctgaa
      721 ggggagctgc ttatccgctt cttcaactcc ttcaatgagg ccaccatggt tctggtctcc
      781 tggatcatgt ggtacgcccc tgtgggcatc atgttcctgg tggctggcaa gatcgtggag
      841 atggaggatg tgggtttact ctttgcccgc cttggcaagt acattctgtg ctgcctgctg
      901 ggtcacgcca tccatgggct cctggtactg cccctcatct acttcctctt cacccgcaaa
      961 aacccctacc gcttcctgtg gggcatcgtg acgccgctgg ccactgcctt tgggacctct
     1021 tccagttccg ccacgctgcc gctgatgatg aagtgcgtgg aggagaataa tggcgtggcc
     1081 aagcacatca gccgtttcat cctgcccatc ggcgccaccg tcaacatgga cggtgccgcg
     1141 ctcttccagt gcgtggccgc agtgttcatt gcacagctca gccagcagtc cttggacttc
     1201 gtaaagatca tcaccatcct ggtcacggcc acagcgtcca gcgtgggggc agcgggcatc
     1261 cctgctggag gtgtcctcac tctggccatc atcctcgaag cagtcaacct cccggtcgac
     1321 catatctcct tgatcctggc tgtggactgg ctagtcgacc ggtcctgtac cgtcctcaat
     1381 gtagaaggtg acgctctggg ggcaggactc ctccaaaatt acgtggaccg tacggagtcg
     1441 agaagcacag agcctgagtt gatacaagtg aagagtgagc tgcccctgga tccgctgcca
     1501 gtccccactg aggaaggaaa ccccctcctc aaacactatc gggggcccgc aggggatgcc
     1561 acggtcgcct ctgagaagga atcagtcatg actagtaccg gtaccggagg tggtggatcc
     1621 gtgagcaagg gcgccgagct gttcaccggc atcgtgccca tcctgatcga gctgaatggc
     1681 gatgtgaatg gccacaagtt cagcgtgagc ggcgagggcg agggcgatgc cacctacggc
     1741 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ctgtgccctg gcccaccctg
     1801 gtgaccaccc tgagctacgg cgtgcagtgc ttctcacgct accccgatca catgaagcag
     1861 cacgacttct tcaagagcgc catgcctgag ggctacatcc aggagcgcac catcttcttc
     1921 gaggatgacg gcaactacaa gtcgcgcgcc gaggtgaagt tcgagggcga taccctggtg
     1981 aatcgcatcg agctgaccgg caccgatttc aaggaggatg gcaacatcct gggcaataag
     2041 atggagtaca actacaacgc ccacaatgtg tacatcatga ccgacaaggc caagaatggc
     2101 atcaaggtga acttcaagat ccgccacaac atcgaggatg gcagcgtgca gctggccgac
     2161 cactaccagc agaatacccc catcggcgat ggccctgtgc tgctgcccga taaccactac
     2221 ctgtccaccc agagcgccct gtccaaggac cccaacgaga agcgcgatca catgatctac
     2281 ttcggcttcg tgaccgccgc cgccatcacc cacggcatgg atgagctgta caagtgagcg
     2341 gccgcgactc tagacccctc tccctccccc ccccctaacg ttactggccg aagccgcttg
     2401 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     2461 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     2521 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     2581 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     2641 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     2701 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     2761 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     2821 ctggggcctc ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     2881 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac
     2941 catggcggaa ggatccgtcg ccaggcagcc tgacctcttg acctgcgacg atgagccgat
     3001 ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg ccgacatgac
     3061 gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg gcgccctgat
     3121 cggccgctct gcggccgatg tcttcgactc ggagacgcac aaccgtctga cgatcgcctt
     3181 ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga tgcgaaagga
     3241 cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg agctcgagcc
     3301 tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca gcgccatccg
     3361 ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc aagaggtgcg
     3421 gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact tcagcggcga
     3481 agtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc actatcctgc
     3541 ctcaaccgtg ccggcgcagg cccgtcggct ctataccatc aacccggtac ggatcattcc
     3601 cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca ccgggcggcc
     3661 gattgatctt agcttcgcca tcctgcgcag cgtctcgccc gtccatctgg agttcatgcg
     3721 caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc gactgtgggg
     3781 attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc gccaagcctg
     3841 cgagctagtc gcccaggttc tggcctggca gatcggcgtg atggaagagt gacgcgtatc
     3901 tgaggggact agggtgtgtt taggcgaaaa gcggggcttc ggttgtacgc ggttaggagt
     3961 cccctcgagg tatggcagat atgcttttgc atagggaggg ggaaatgtag tcttaatcat
     4021 aggttaacat gtatattacc aaataaggga atcgcctgat gcaccaaata aggtattata
     4081 tgatcccatt ggtggcgagg gagcgacctg agggcatatg ggcgttaaca gaactgtctg
     4141 tccttgcgtc attccttcct catcggatca tgtacgcggc agagtatgat tggataacag
     4201 gatggcacca ttcatcgtgg cgcatgctga ttggtgcgac taaggagtta tgtaacccac
     4261 gaatgtactt aagcttgtag ttgctaacaa taaagggcca ttctacctct caccacattg
     4321 gtgtgcacct gggttgatgg ccggaccgtc gattccctga cgactacgag cacctgaatg
     4381 aagcagaagg catcctgcag gaattcctcg agacgtacgg tgcgcgcgat gcattcgaag
     4441 atctgcccta tagtgagtcg tattacgtag cttggcgtaa tcatggtcat agctgtttcc
     4501 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4561 taaagcctgg ggtgcctaat gagtgagcta actcacatta catgtgagca aaaggccagc
     4621 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4681 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4741 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4801 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4861 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4921 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4981 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5041 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5101 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5161 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5221 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5281 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5341 tcttcaccta gatcctttta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5401 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5461 ttcccttcct ttctcgccac gttcgctttc cccgtcaagc tctaaatcgg gggctccctt
     5521 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     5581 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5641 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     5701 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5761 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt taaatatttg
     5821 cttatacaat cttcctgttt ttggggcttt tctgattatc aaccggggta aatcaatcta
     5881 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5941 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6001 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6061 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6121 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6181 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6241 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6301 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6361 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6421 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6481 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     6541 cgcgccacat agcagaactt taaaagtgct catcattgga gaacgttctt cggggcgaaa
     6601 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6661 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6721 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6781 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6841 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6901 tgacgtagtt aacaaaaaaa agcccgccga agcgggcttt attaccaagc gaagcgccat
     6961 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     7021 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     7081 tcccagtcac gacgttgtaa aacgacggcc agtccgtaat acgactcact ataggtgtag
     7141 tcttaatcat aggttaacat gtatattacc aaataaggga atcgcctgat gcaccaaata
     7201 aggtattata tgatcccatt ggtggcgagg gagcgacctg agggcatatg ggcgttaaca
     7261 gaactgtctg tccttgcgtc attccttcct catcggatca tgtacgcggc agagtatgat
     7321 tggataacag gatggcacca ttcatcgtgg cgcatgctga ttggtgcgac taaggagtta
     7381 tgtaacccac gaatgtactt aagcttgtag ttgctaacaa taaagggcca ttctacctct
     7441 caccacattg gtgtgcacct gggttgatgg ccggaccgtc gattccctga cgactacgag
     7501 cacctgaatg aagcagaagg cttcatttgg tgaccccgac gtgatcgtta gggaatagtg
     7561 gtcggccaca ggcggcgtgg cgatcctgtc tccatccgtc tcgtcgaccg ggaggcgagt
     7621 tcgatgaccc tgatagaggg ggctgcggct tagggaggca gaagctgagc accgtcggta
     7681 ggagctccag ggcccggagc gactgacccc tgccgagaac tcagagggtc gtcggaagac
     7741 ggagagtgag cccgacgacc accccaggca cgtctttggt cggcctgcgg atcaagctag
     7801 caagccgtca ttaaggtgat ttcgtccgcg tgtaaaacct attgcgggaa aatctctcct
     7861 tctaagaagg aaataggggc catgttgtcc ctgttacaaa aggaagggtt gctttagtct
     7921 ccctcagatt tatattctcc ggggtcctgg gatcccatca ctgcggcgct ctcccagcgg
     7981 gcaatggtac ttggaaaatc gggagagtta aaaacctggg gattggtttt gggggcattg
     8041 aaggcggctc gagagtaaca ggactaactt gaatcgatgc gctaccggac tcagatcttg
     8101 ccacc
//
