LOCUS       Exported                8138 bp ds-DNA     circular SYN 05-FEB-2021
DEFINITION  pFuTraP vector expressing human ASCT2 with N212Q substitution.
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..8138
                     /organism="recombinant plasmid"
                     /lab_host="Gallus gallus"
                     /mol_type="other DNA"
     CDS             1..1623
                     /codon_start=1
                     /product="ASCT2"
                     /label=hASCT2
                     /note="human ASCT2"
                     /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV
                     RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII
                     LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN
                     ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSYSTTYEERQITGTRVKVPV
                     GQEVEGMNILGLVVFAIVFGVALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGI
                     MFLVAGKIVEMEDVGLLFARLGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGI
                     VTPLATAFGTSSSSATLPLMMKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAV
                     FIAQLSQQSLDFVKIITILVTATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILA
                     VDWLVDRSCTVLNVEGDALGAGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEG
                     NPLLKHYRGPAGDATVASEKESVM"
     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
     mat_peptide     610..681
                     /label=region C
     misc_feature    694..744
                     /label=TM4c
                     /note="Garaeva et al., 2018"
     misc_feature    760..876
                     /label=TM5
                     /note="Garaeva et al., 2018"
     misc_feature    889..987
                     /label=TM6
                     /note="Garaeva et al., 2018"
     misc_feature    997..1050
                     /label=HP1a
                     /note="Garaeva et al., 2018"
     misc_feature    1057..1098
                     /label=HP1b
                     /note="Garaeva et al., 2018"
     misc_feature    1114..1155
                     /label=TM7a
                     /note="Garaeva et al., 2018"
     misc_feature    1162..1209
                     /label=TM7b
                     /note="Garaeva et al., 2018"
     misc_feature    1231..1275
                     /label=HP2a
                     /note="Garaeva et al., 2018"
     misc_feature    1300..1338
                     /label=HP2b
                     /note="Garaeva et al., 2018"
     misc_feature    1363..1482
                     /label=TM8
                     /note="Garaeva et al., 2018"
     CDS             1639..1653
                     /codon_start=1
                     /label=GGGGS linker
                     /translation="GGGGS"
     CDS             1654..2370
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     misc_feature    2388..2974
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2975..3925
                     /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             4039..4424
                     /label=dLTR
     V_segment       complement(4425..7168)
                     /label=pLitmus 28i
     promoter        complement(4480..4498)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4519..4535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4543..4559
                     /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(4567..4597)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4612..4633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4694..5282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5393..5902
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5944..6804)
                     /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(6805..6909)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7130..7146
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7150..7168
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             7169..7560
     misc_feature    7561..7578
                     /label=PBS
                     /note="the same as for RCAS"
     misc_feature    7615..7719
                     /label=packaging signal
                     /note="according to Banks JD and Linial ML, J. Virol., 
                     2000"
     gene            7831..8093
                     /label=p19
                     /note="p19"
     gene            8094..8107
                     /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 aggcagatca ccggaaccag ggtgaaggtg
      661 cccgtggggc aggaggtgga ggggatgaac atcctgggct tggtagtgtt tgcaattgtc
      721 tttggtgtgg cgctgcggaa gctggggcct gaaggggagc tgcttatccg cttcttcaac
      781 tccttcaatg aggccaccat ggttctggtc tcctggatca tgtggtacgc ccctgtgggc
      841 atcatgttcc tggtggctgg caagatcgtg gagatggagg atgtgggttt actctttgcc
      901 cgccttggca agtacattct gtgctgcctg ctgggtcacg ccatccatgg gctcctggta
      961 ctgcccctca tctacttcct cttcacccgc aaaaacccct accgcttcct gtggggcatc
     1021 gtgacgccgc tggccactgc ctttgggacc tcttccagtt ccgccacgct gccgctgatg
     1081 atgaagtgcg tggaggagaa taatggcgtg gccaagcaca tcagccgttt catcctgccc
     1141 atcggcgcca ccgtcaacat ggacggtgcc gcgctcttcc agtgcgtggc cgcagtgttc
     1201 attgcacagc tcagccagca gtccttggac ttcgtaaaga tcatcaccat cctggtcacg
     1261 gccacagcgt ccagcgtggg ggcagcgggc atccctgctg gaggtgtcct cactctggcc
     1321 atcatcctcg aagcagtcaa cctcccggtc gaccatatct ccttgatcct ggctgtggac
     1381 tggctagtcg accggtcctg taccgtcctc aatgtagaag gtgacgctct gggggcagga
     1441 ctcctccaaa attacgtgga ccgtacggag tcgagaagca cagagcctga gttgatacaa
     1501 gtgaagagtg agctgcccct ggatccgctg ccagtcccca ctgaggaagg aaaccccctc
     1561 ctcaaacact atcgggggcc cgcaggggat gccacggtcg cctctgagaa ggaatcagtc
     1621 atgactagta ccggtaccgg aggtggtgga tccgtgagca agggcgccga gctgttcacc
     1681 ggcatcgtgc ccatcctgat cgagctgaat ggcgatgtga atggccacaa gttcagcgtg
     1741 agcggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1801 accggcaagc tgcctgtgcc ctggcccacc ctggtgacca ccctgagcta cggcgtgcag
     1861 tgcttctcac gctaccccga tcacatgaag cagcacgact tcttcaagag cgccatgcct
     1921 gagggctaca tccaggagcg caccatcttc ttcgaggatg acggcaacta caagtcgcgc
     1981 gccgaggtga agttcgaggg cgataccctg gtgaatcgca tcgagctgac cggcaccgat
     2041 ttcaaggagg atggcaacat cctgggcaat aagatggagt acaactacaa cgcccacaat
     2101 gtgtacatca tgaccgacaa ggccaagaat ggcatcaagg tgaacttcaa gatccgccac
     2161 aacatcgagg atggcagcgt gcagctggcc gaccactacc agcagaatac ccccatcggc
     2221 gatggccctg tgctgctgcc cgataaccac tacctgtcca cccagagcgc cctgtccaag
     2281 gaccccaacg agaagcgcga tcacatgatc tacttcggct tcgtgaccgc cgccgccatc
     2341 acccacggca tggatgagct gtacaagtga gcggccgcga ctctagaccc ctctccctcc
     2401 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     2461 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     2521 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     2581 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     2641 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     2701 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     2761 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2821 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     2881 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2941 ctttgaaaaa cacgatgata atatggccac aaccatggcg gaaggatccg tcgccaggca
     3001 gcctgacctc ttgacctgcg acgatgagcc gatccatatc cccggtgcca tccaaccgca
     3061 tggactgctg ctcgccctcg ccgccgacat gacgatcgtt gccggcagcg acaaccttcc
     3121 cgaactcacc ggactggcga tcggcgccct gatcggccgc tctgcggccg atgtcttcga
     3181 ctcggagacg cacaaccgtc tgacgatcgc cttggccgag cccggggcgg ccgtcggagc
     3241 accgatcact gtcggcttca cgatgcgaaa ggacgcaggc ttcatcggct cctggcatcg
     3301 ccatgatcag ctcatcttcc tcgagctcga gcctccccag cgggacgtcg ccgagccgca
     3361 ggcgttcttc cgccgcacca acagcgccat ccgccgcctg caggccgccg aaaccttgga
     3421 aagcgcctgc gccgccgcgg cgcaagaggt gcggaagatt accggcttcg atcgggtgat
     3481 gatctatcgc ttcgcctccg acttcagcgg cgaagtgatc gcagaggatc ggtgcgccga
     3541 ggtcgagtca aaactaggcc tgcactatcc tgcctcaacc gtgccggcgc aggcccgtcg
     3601 gctctatacc atcaacccgg tacggatcat tcccgatatc aattatcggc cggtgccggt
     3661 caccccagac ctcaatccgg tcaccgggcg gccgattgat cttagcttcg ccatcctgcg
     3721 cagcgtctcg cccgtccatc tggagttcat gcgcaacata ggcatgcacg gcacgatgtc
     3781 gatctcgatt ttgcgcggcg agcgactgtg gggattgatc gtttgccatc accgaacgcc
     3841 gtactacgtc gatctcgatg gccgccaagc ctgcgagcta gtcgcccagg ttctggcctg
     3901 gcagatcggc gtgatggaag agtgacgcgt atctgagggg actagggtgt gtttaggcga
     3961 aaagcggggc ttcggttgta cgcggttagg agtcccctcg aggtatggca gatatgcttt
     4021 tgcataggga gggggaaatg tagtcttaat cataggttaa catgtatatt accaaataag
     4081 ggaatcgcct gatgcaccaa ataaggtatt atatgatccc attggtggcg agggagcgac
     4141 ctgagggcat atgggcgtta acagaactgt ctgtccttgc gtcattcctt cctcatcgga
     4201 tcatgtacgc ggcagagtat gattggataa caggatggca ccattcatcg tggcgcatgc
     4261 tgattggtgc gactaaggag ttatgtaacc cacgaatgta cttaagcttg tagttgctaa
     4321 caataaaggg ccattctacc tctcaccaca ttggtgtgca cctgggttga tggccggacc
     4381 gtcgattccc tgacgactac gagcacctga atgaagcaga aggcatcctg caggaattcc
     4441 tcgagacgta cggtgcgcgc gatgcattcg aagatctgcc ctatagtgag tcgtattacg
     4501 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4561 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4621 ctaactcaca ttacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4681 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4741 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4801 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4861 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4921 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4981 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5041 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5101 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5161 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5221 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5281 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5341 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttacgcgccc
     5401 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5461 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgct
     5521 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5581 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     5641 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5701 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     5761 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5821 acaaaatatt aacgtttaca atttaaatat ttgcttatac aatcttcctg tttttggggc
     5881 ttttctgatt atcaaccggg gtaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5941 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6001 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6061 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6121 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6181 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6241 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6301 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6361 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6421 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6481 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6541 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     6601 gctcatcatt ggagaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6661 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6721 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6781 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6841 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6901 ggttccgcgc acatttcccc gaaaagtgcc acctgacgta gttaacaaaa aaaagcccgc
     6961 cgaagcgggc tttattacca agcgaagcgc cattcgccat tcaggctgcg caactgttgg
     7021 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     7081 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     7141 gccagtccgt aatacgactc actataggtg tagtcttaat cataggttaa catgtatatt
     7201 accaaataag ggaatcgcct gatgcaccaa ataaggtatt atatgatccc attggtggcg
     7261 agggagcgac ctgagggcat atgggcgtta acagaactgt ctgtccttgc gtcattcctt
     7321 cctcatcgga tcatgtacgc ggcagagtat gattggataa caggatggca ccattcatcg
     7381 tggcgcatgc tgattggtgc gactaaggag ttatgtaacc cacgaatgta cttaagcttg
     7441 tagttgctaa caataaaggg ccattctacc tctcaccaca ttggtgtgca cctgggttga
     7501 tggccggacc gtcgattccc tgacgactac gagcacctga atgaagcaga aggcttcatt
     7561 tggtgacccc gacgtgatcg ttagggaata gtggtcggcc acaggcggcg tggcgatcct
     7621 gtctccatcc gtctcgtcga ccgggaggcg agttcgatga ccctgataga gggggctgcg
     7681 gcttagggag gcagaagctg agcaccgtcg gtaggagctc cagggcccgg agcgactgac
     7741 ccctgccgag aactcagagg gtcgtcggaa gacggagagt gagcccgacg accaccccag
     7801 gcacgtcttt ggtcggcctg cggatcaagc tagcaagccg tcattaaggt gatttcgtcc
     7861 gcgtgtaaaa cctattgcgg gaaaatctct ccttctaaga aggaaatagg ggccatgttg
     7921 tccctgttac aaaaggaagg gttgctttag tctccctcag atttatattc tccggggtcc
     7981 tgggatccca tcactgcggc gctctcccag cgggcaatgg tacttggaaa atcgggagag
     8041 ttaaaaacct ggggattggt tttgggggca ttgaaggcgg ctcgagagta acaggactaa
     8101 cttgaatcga tgcgctaccg gactcagatc ttgccacc
//
