LOCUS       Exported                8111 bp ds-DNA     circular SYN 05-FEB-2021
DEFINITION  pFuTraP vector expressing human ASCT2 with d11 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..8111
                     /organism="recombinant plasmid"
                     /lab_host="Gallus gallus"
                     /mol_type="other DNA"
     CDS             1..1596
                     /codon_start=1
                     /label=hASCT2-d11
                     /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV
                     RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII
                     LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN
                     ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYSTTYGGVPVGQEVEGMNI
                     LGLVVFAIVFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFLVAGKIV
                     EMEDVGLLFARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTPLATAFG
                     TSSSSATLPLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIAQLSQQS
                     LDFVKIITILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDWLVDRSC
                     TVLNVEGDALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPLLKHYRG
                     PAGDATVASEKESVM"
     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    625..630
                     /label=d11
     misc_feature    667..717
                     /label=TM4c
                     /note="Garaeva et al., 2018"
     misc_feature    733..849
                     /label=TM5
                     /note="Garaeva et al., 2018"
     misc_feature    862..960
                     /label=TM6
                     /note="Garaeva et al., 2018"
     misc_feature    970..1023
                     /label=HP1a
                     /note="Garaeva et al., 2018"
     misc_feature    1030..1071
                     /label=HP1b
                     /note="Garaeva et al., 2018"
     misc_feature    1087..1128
                     /label=TM7a
                     /note="Garaeva et al., 2018"
     misc_feature    1135..1182
                     /label=TM7b
                     /note="Garaeva et al., 2018"
     misc_feature    1204..1248
                     /label=HP2a
                     /note="Garaeva et al., 2018"
     misc_feature    1273..1311
                     /label=HP2b
                     /note="Garaeva et al., 2018"
     misc_feature    1336..1455
                     /label=TM8
                     /note="Garaeva et al., 2018"
     CDS             1612..1626
                     /codon_start=1
                     /label=GGGGS linker
                     /translation="GGGGS"
     CDS             1627..2343
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     misc_feature    2361..2947
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2948..3898
                     /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             4012..4397
                     /label=dLTR
     V_segment       complement(4398..7141)
                     /label=pLitmus 28i
     promoter        complement(4453..4471)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4492..4508)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4516..4532
                     /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(4540..4570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4585..4606
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4667..5255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5366..5875
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5917..6777)
                     /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(6778..6882)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7103..7119
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7123..7141
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             7142..7533
     misc_feature    7534..7551
                     /label=PBS
                     /note="the same as for RCAS"
     misc_feature    7588..7692
                     /label=packaging signal
                     /note="according to Banks JD and Linial ML, J. Virol., 
                     2000"
     gene            7804..8066
                     /label=p19
                     /note="p19"
     gene            8067..8080
                     /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 ctatggaggg gtacccgtgg ggcaggaggt ggaggggatg
      661 aacatcctgg gcttggtagt gtttgcaatt gtctttggtg tggcgctgcg gaagctgggg
      721 cctgaagggg agctgcttat ccgcttcttc aactccttca atgaggccac catggttctg
      781 gtctcctgga tcatgtggta cgcccctgtg ggcatcatgt tcctggtggc tggcaagatc
      841 gtggagatgg aggatgtggg tttactcttt gcccgccttg gcaagtacat tctgtgctgc
      901 ctgctgggtc acgccatcca tgggctcctg gtactgcccc tcatctactt cctcttcacc
      961 cgcaaaaacc cctaccgctt cctgtggggc atcgtgacgc cgctggccac tgcctttggg
     1021 acctcttcca gttccgccac gctgccgctg atgatgaagt gcgtggagga gaataatggc
     1081 gtggccaagc acatcagccg tttcatcctg cccatcggcg ccaccgtcaa catggacggt
     1141 gccgcgctct tccagtgcgt ggccgcagtg ttcattgcac agctcagcca gcagtccttg
     1201 gacttcgtaa agatcatcac catcctggtc acggccacag cgtccagcgt gggggcagcg
     1261 ggcatccctg ctggaggtgt cctcactctg gccatcatcc tcgaagcagt caacctcccg
     1321 gtcgaccata tctccttgat cctggctgtg gactggctag tcgaccggtc ctgtaccgtc
     1381 ctcaatgtag aaggtgacgc tctgggggca ggactcctcc aaaattacgt ggaccgtacg
     1441 gagtcgagaa gcacagagcc tgagttgata caagtgaaga gtgagctgcc cctggatccg
     1501 ctgccagtcc ccactgagga aggaaacccc ctcctcaaac actatcgggg gcccgcaggg
     1561 gatgccacgg tcgcctctga gaaggaatca gtcatgacta gtaccggtac cggaggtggt
     1621 ggatccgtga gcaagggcgc cgagctgttc accggcatcg tgcccatcct gatcgagctg
     1681 aatggcgatg tgaatggcca caagttcagc gtgagcggcg agggcgaggg cgatgccacc
     1741 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcctgt gccctggccc
     1801 accctggtga ccaccctgag ctacggcgtg cagtgcttct cacgctaccc cgatcacatg
     1861 aagcagcacg acttcttcaa gagcgccatg cctgagggct acatccagga gcgcaccatc
     1921 ttcttcgagg atgacggcaa ctacaagtcg cgcgccgagg tgaagttcga gggcgatacc
     1981 ctggtgaatc gcatcgagct gaccggcacc gatttcaagg aggatggcaa catcctgggc
     2041 aataagatgg agtacaacta caacgcccac aatgtgtaca tcatgaccga caaggccaag
     2101 aatggcatca aggtgaactt caagatccgc cacaacatcg aggatggcag cgtgcagctg
     2161 gccgaccact accagcagaa tacccccatc ggcgatggcc ctgtgctgct gcccgataac
     2221 cactacctgt ccacccagag cgccctgtcc aaggacccca acgagaagcg cgatcacatg
     2281 atctacttcg gcttcgtgac cgccgccgcc atcacccacg gcatggatga gctgtacaag
     2341 tgagcggccg cgactctaga cccctctccc tccccccccc ctaacgttac tggccgaagc
     2401 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     2461 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     2521 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     2581 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     2641 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     2701 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     2761 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     2821 tctgatctgg ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     2881 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     2941 cacaaccatg gcggaaggat ccgtcgccag gcagcctgac ctcttgacct gcgacgatga
     3001 gccgatccat atccccggtg ccatccaacc gcatggactg ctgctcgccc tcgccgccga
     3061 catgacgatc gttgccggca gcgacaacct tcccgaactc accggactgg cgatcggcgc
     3121 cctgatcggc cgctctgcgg ccgatgtctt cgactcggag acgcacaacc gtctgacgat
     3181 cgccttggcc gagcccgggg cggccgtcgg agcaccgatc actgtcggct tcacgatgcg
     3241 aaaggacgca ggcttcatcg gctcctggca tcgccatgat cagctcatct tcctcgagct
     3301 cgagcctccc cagcgggacg tcgccgagcc gcaggcgttc ttccgccgca ccaacagcgc
     3361 catccgccgc ctgcaggccg ccgaaacctt ggaaagcgcc tgcgccgccg cggcgcaaga
     3421 ggtgcggaag attaccggct tcgatcgggt gatgatctat cgcttcgcct ccgacttcag
     3481 cggcgaagtg atcgcagagg atcggtgcgc cgaggtcgag tcaaaactag gcctgcacta
     3541 tcctgcctca accgtgccgg cgcaggcccg tcggctctat accatcaacc cggtacggat
     3601 cattcccgat atcaattatc ggccggtgcc ggtcacccca gacctcaatc cggtcaccgg
     3661 gcggccgatt gatcttagct tcgccatcct gcgcagcgtc tcgcccgtcc atctggagtt
     3721 catgcgcaac ataggcatgc acggcacgat gtcgatctcg attttgcgcg gcgagcgact
     3781 gtggggattg atcgtttgcc atcaccgaac gccgtactac gtcgatctcg atggccgcca
     3841 agcctgcgag ctagtcgccc aggttctggc ctggcagatc ggcgtgatgg aagagtgacg
     3901 cgtatctgag gggactaggg tgtgtttagg cgaaaagcgg ggcttcggtt gtacgcggtt
     3961 aggagtcccc tcgaggtatg gcagatatgc ttttgcatag ggagggggaa atgtagtctt
     4021 aatcataggt taacatgtat attaccaaat aagggaatcg cctgatgcac caaataaggt
     4081 attatatgat cccattggtg gcgagggagc gacctgaggg catatgggcg ttaacagaac
     4141 tgtctgtcct tgcgtcattc cttcctcatc ggatcatgta cgcggcagag tatgattgga
     4201 taacaggatg gcaccattca tcgtggcgca tgctgattgg tgcgactaag gagttatgta
     4261 acccacgaat gtacttaagc ttgtagttgc taacaataaa gggccattct acctctcacc
     4321 acattggtgt gcacctgggt tgatggccgg accgtcgatt ccctgacgac tacgagcacc
     4381 tgaatgaagc agaaggcatc ctgcaggaat tcctcgagac gtacggtgcg cgcgatgcat
     4441 tcgaagatct gccctatagt gagtcgtatt acgtagcttg gcgtaatcat ggtcatagct
     4501 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4561 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattacatg tgagcaaaag
     4621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5341 aaaggatctt cacctagatc cttttacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5401 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5461 gctttcttcc cttcctttct cgccacgttc gctttccccg tcaagctcta aatcgggggc
     5521 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     5581 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5641 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5701 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5761 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa
     5821 tatttgctta tacaatcttc ctgtttttgg ggcttttctg attatcaacc ggggtaaatc
     5881 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5941 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6001 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6061 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6121 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6181 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6241 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6301 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6361 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6421 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6481 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga
     6541 taataccgcg ccacatagca gaactttaaa agtgctcatc attggagaac gttcttcggg
     6601 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6661 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6721 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6781 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6841 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6901 gccacctgac gtagttaaca aaaaaaagcc cgccgaagcg ggctttatta ccaagcgaag
     6961 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7021 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     7081 gggttttccc agtcacgacg ttgtaaaacg acggccagtc cgtaatacga ctcactatag
     7141 gtgtagtctt aatcataggt taacatgtat attaccaaat aagggaatcg cctgatgcac
     7201 caaataaggt attatatgat cccattggtg gcgagggagc gacctgaggg catatgggcg
     7261 ttaacagaac tgtctgtcct tgcgtcattc cttcctcatc ggatcatgta cgcggcagag
     7321 tatgattgga taacaggatg gcaccattca tcgtggcgca tgctgattgg tgcgactaag
     7381 gagttatgta acccacgaat gtacttaagc ttgtagttgc taacaataaa gggccattct
     7441 acctctcacc acattggtgt gcacctgggt tgatggccgg accgtcgatt ccctgacgac
     7501 tacgagcacc tgaatgaagc agaaggcttc atttggtgac cccgacgtga tcgttaggga
     7561 atagtggtcg gccacaggcg gcgtggcgat cctgtctcca tccgtctcgt cgaccgggag
     7621 gcgagttcga tgaccctgat agagggggct gcggcttagg gaggcagaag ctgagcaccg
     7681 tcggtaggag ctccagggcc cggagcgact gacccctgcc gagaactcag agggtcgtcg
     7741 gaagacggag agtgagcccg acgaccaccc caggcacgtc tttggtcggc ctgcggatca
     7801 agctagcaag ccgtcattaa ggtgatttcg tccgcgtgta aaacctattg cgggaaaatc
     7861 tctccttcta agaaggaaat aggggccatg ttgtccctgt tacaaaagga agggttgctt
     7921 tagtctccct cagatttata ttctccgggg tcctgggatc ccatcactgc ggcgctctcc
     7981 cagcgggcaa tggtacttgg aaaatcggga gagttaaaaa cctggggatt ggttttgggg
     8041 gcattgaagg cggctcgaga gtaacaggac taacttgaat cgatgcgcta ccggactcag
     8101 atcttgccac c
//
