LOCUS       Exported                8078 bp ds-DNA     circular SYN 05-FEB-2021
DEFINITION  pFuTraP vector expressing human ASCT2 with d22 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..8078
                     /organism="recombinant plasmid"
                     /lab_host="Gallus gallus"
                     /mol_type="other DNA"
     CDS             1..1563
                     /codon_start=1
                     /label=hASCT2-d22
                     /translation="MVADPPRDSKGLAAAEPTANGGLALASIEDQGAAAGGYCGSRDQV
                     RRCLRANLLVLLTVVAVVAGVALGLGVSGAGGALALGPERLSAFVFPGELLLRLLRMII
                     LPLVVCSLIGGAASLDPGALGRLGAWALLFFLVTTLLASALGVGLALALQPGAASAAIN
                     ASVGAAGSAENAPSKEVLDSFLDLARNIFPSNLVSAAFRSGGVEGMNILGLVVFAIVFG
                     VALRKLGPEGELLIRFFNSFNEATMVLVSWIMWYAPVGIMFLVAGKIVEMEDVGLLFAR
                     LGKYILCCLLGHAIHGLLVLPLIYFLFTRKNPYRFLWGIVTPLATAFGTSSSSATLPLM
                     MKCVEENNGVAKHISRFILPIGATVNMDGAALFQCVAAVFIAQLSQQSLDFVKIITILV
                     TATASSVGAAGIPAGGVLTLAIILEAVNLPVDHISLILAVDWLVDRSCTVLNVEGDALG
                     AGLLQNYVDRTESRSTEPELIQVKSELPLDPLPVPTEEGNPLLKHYRGPAGDATVASEK
                     ESVM"
     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    610..615
                     /label=d22
     misc_feature    634..684
                     /label=TM4c
                     /note="Garaeva et al., 2018"
     misc_feature    700..816
                     /label=TM5
                     /note="Garaeva et al., 2018"
     misc_feature    829..927
                     /label=TM6
                     /note="Garaeva et al., 2018"
     misc_feature    937..990
                     /label=HP1a
                     /note="Garaeva et al., 2018"
     misc_feature    997..1038
                     /label=HP1b
                     /note="Garaeva et al., 2018"
     misc_feature    1054..1095
                     /label=TM7a
                     /note="Garaeva et al., 2018"
     misc_feature    1102..1149
                     /label=TM7b
                     /note="Garaeva et al., 2018"
     misc_feature    1171..1215
                     /label=HP2a
                     /note="Garaeva et al., 2018"
     misc_feature    1240..1278
                     /label=HP2b
                     /note="Garaeva et al., 2018"
     misc_feature    1303..1422
                     /label=TM8
                     /note="Garaeva et al., 2018"
     CDS             1579..1593
                     /codon_start=1
                     /label=GGGGS linker
                     /translation="GGGGS"
     CDS             1594..2310
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     misc_feature    2328..2914
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2915..3865
                     /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             3979..4364
                     /label=dLTR
     V_segment       complement(4365..7108)
                     /label=pLitmus 28i
     promoter        complement(4420..4438)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4420..4438)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4459..4475)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4483..4499
                     /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(4507..4537)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4552..4573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4634..5222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5333..5842
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(5884..6744)
                     /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(6745..6849)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7070..7086
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7090..7108
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7090..7108
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             7109..7500
     LTR             7109..7494
                     /label=dLTR
     misc_feature    7501..7518
                     /label=PBS
                     /note="the same as for RCAS"
     misc_feature    7555..7659
                     /label=packaging signal
                     /note="according to Banks JD and Linial ML, J. Virol., 
                     2000"
     gene            7771..8033
                     /label=p19
                     /note="p19"
     gene            8034..8047
                     /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 tttcgctccg gaggagtcga ggggatgaac atcctgggct tggtagtgtt tgcaattgtc
      661 tttggtgtgg cgctgcggaa gctggggcct gaaggggagc tgcttatccg cttcttcaac
      721 tccttcaatg aggccaccat ggttctggtc tcctggatca tgtggtacgc ccctgtgggc
      781 atcatgttcc tggtggctgg caagatcgtg gagatggagg atgtgggttt actctttgcc
      841 cgccttggca agtacattct gtgctgcctg ctgggtcacg ccatccatgg gctcctggta
      901 ctgcccctca tctacttcct cttcacccgc aaaaacccct accgcttcct gtggggcatc
      961 gtgacgccgc tggccactgc ctttgggacc tcttccagtt ccgccacgct gccgctgatg
     1021 atgaagtgcg tggaggagaa taatggcgtg gccaagcaca tcagccgttt catcctgccc
     1081 atcggcgcca ccgtcaacat ggacggtgcc gcgctcttcc agtgcgtggc cgcagtgttc
     1141 attgcacagc tcagccagca gtccttggac ttcgtaaaga tcatcaccat cctggtcacg
     1201 gccacagcgt ccagcgtggg ggcagcgggc atccctgctg gaggtgtcct cactctggcc
     1261 atcatcctcg aagcagtcaa cctcccggtc gaccatatct ccttgatcct ggctgtggac
     1321 tggctagtcg accggtcctg taccgtcctc aatgtagaag gtgacgctct gggggcagga
     1381 ctcctccaaa attacgtgga ccgtacggag tcgagaagca cagagcctga gttgatacaa
     1441 gtgaagagtg agctgcccct ggatccgctg ccagtcccca ctgaggaagg aaaccccctc
     1501 ctcaaacact atcgggggcc cgcaggggat gccacggtcg cctctgagaa ggaatcagtc
     1561 atgactagta ccggtaccgg aggtggtgga tccgtgagca agggcgccga gctgttcacc
     1621 ggcatcgtgc ccatcctgat cgagctgaat ggcgatgtga atggccacaa gttcagcgtg
     1681 agcggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1741 accggcaagc tgcctgtgcc ctggcccacc ctggtgacca ccctgagcta cggcgtgcag
     1801 tgcttctcac gctaccccga tcacatgaag cagcacgact tcttcaagag cgccatgcct
     1861 gagggctaca tccaggagcg caccatcttc ttcgaggatg acggcaacta caagtcgcgc
     1921 gccgaggtga agttcgaggg cgataccctg gtgaatcgca tcgagctgac cggcaccgat
     1981 ttcaaggagg atggcaacat cctgggcaat aagatggagt acaactacaa cgcccacaat
     2041 gtgtacatca tgaccgacaa ggccaagaat ggcatcaagg tgaacttcaa gatccgccac
     2101 aacatcgagg atggcagcgt gcagctggcc gaccactacc agcagaatac ccccatcggc
     2161 gatggccctg tgctgctgcc cgataaccac tacctgtcca cccagagcgc cctgtccaag
     2221 gaccccaacg agaagcgcga tcacatgatc tacttcggct tcgtgaccgc cgccgccatc
     2281 acccacggca tggatgagct gtacaagtga gcggccgcga ctctagaccc ctctccctcc
     2341 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     2401 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     2461 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     2521 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     2581 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     2641 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     2701 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     2761 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     2821 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     2881 ctttgaaaaa cacgatgata atatggccac aaccatggcg gaaggatccg tcgccaggca
     2941 gcctgacctc ttgacctgcg acgatgagcc gatccatatc cccggtgcca tccaaccgca
     3001 tggactgctg ctcgccctcg ccgccgacat gacgatcgtt gccggcagcg acaaccttcc
     3061 cgaactcacc ggactggcga tcggcgccct gatcggccgc tctgcggccg atgtcttcga
     3121 ctcggagacg cacaaccgtc tgacgatcgc cttggccgag cccggggcgg ccgtcggagc
     3181 accgatcact gtcggcttca cgatgcgaaa ggacgcaggc ttcatcggct cctggcatcg
     3241 ccatgatcag ctcatcttcc tcgagctcga gcctccccag cgggacgtcg ccgagccgca
     3301 ggcgttcttc cgccgcacca acagcgccat ccgccgcctg caggccgccg aaaccttgga
     3361 aagcgcctgc gccgccgcgg cgcaagaggt gcggaagatt accggcttcg atcgggtgat
     3421 gatctatcgc ttcgcctccg acttcagcgg cgaagtgatc gcagaggatc ggtgcgccga
     3481 ggtcgagtca aaactaggcc tgcactatcc tgcctcaacc gtgccggcgc aggcccgtcg
     3541 gctctatacc atcaacccgg tacggatcat tcccgatatc aattatcggc cggtgccggt
     3601 caccccagac ctcaatccgg tcaccgggcg gccgattgat cttagcttcg ccatcctgcg
     3661 cagcgtctcg cccgtccatc tggagttcat gcgcaacata ggcatgcacg gcacgatgtc
     3721 gatctcgatt ttgcgcggcg agcgactgtg gggattgatc gtttgccatc accgaacgcc
     3781 gtactacgtc gatctcgatg gccgccaagc ctgcgagcta gtcgcccagg ttctggcctg
     3841 gcagatcggc gtgatggaag agtgacgcgt atctgagggg actagggtgt gtttaggcga
     3901 aaagcggggc ttcggttgta cgcggttagg agtcccctcg aggtatggca gatatgcttt
     3961 tgcataggga gggggaaatg tagtcttaat cataggttaa catgtatatt accaaataag
     4021 ggaatcgcct gatgcaccaa ataaggtatt atatgatccc attggtggcg agggagcgac
     4081 ctgagggcat atgggcgtta acagaactgt ctgtccttgc gtcattcctt cctcatcgga
     4141 tcatgtacgc ggcagagtat gattggataa caggatggca ccattcatcg tggcgcatgc
     4201 tgattggtgc gactaaggag ttatgtaacc cacgaatgta cttaagcttg tagttgctaa
     4261 caataaaggg ccattctacc tctcaccaca ttggtgtgca cctgggttga tggccggacc
     4321 gtcgattccc tgacgactac gagcacctga atgaagcaga aggcatcctg caggaattcc
     4381 tcgagacgta cggtgcgcgc gatgcattcg aagatctgcc ctatagtgag tcgtattacg
     4441 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4501 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4561 ctaactcaca ttacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4621 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4681 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4741 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4801 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4861 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4921 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4981 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5041 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5101 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5161 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5221 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5281 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttacgcgccc
     5341 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5401 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgct
     5461 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5521 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     5581 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5641 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     5701 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5761 acaaaatatt aacgtttaca atttaaatat ttgcttatac aatcttcctg tttttggggc
     5821 ttttctgatt atcaaccggg gtaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5881 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5941 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6001 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6061 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6121 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6181 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6241 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6301 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6361 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6421 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6481 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     6541 gctcatcatt ggagaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6601 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6661 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6721 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6781 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6841 ggttccgcgc acatttcccc gaaaagtgcc acctgacgta gttaacaaaa aaaagcccgc
     6901 cgaagcgggc tttattacca agcgaagcgc cattcgccat tcaggctgcg caactgttgg
     6961 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     7021 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     7081 gccagtccgt aatacgactc actataggtg tagtcttaat cataggttaa catgtatatt
     7141 accaaataag ggaatcgcct gatgcaccaa ataaggtatt atatgatccc attggtggcg
     7201 agggagcgac ctgagggcat atgggcgtta acagaactgt ctgtccttgc gtcattcctt
     7261 cctcatcgga tcatgtacgc ggcagagtat gattggataa caggatggca ccattcatcg
     7321 tggcgcatgc tgattggtgc gactaaggag ttatgtaacc cacgaatgta cttaagcttg
     7381 tagttgctaa caataaaggg ccattctacc tctcaccaca ttggtgtgca cctgggttga
     7441 tggccggacc gtcgattccc tgacgactac gagcacctga atgaagcaga aggcttcatt
     7501 tggtgacccc gacgtgatcg ttagggaata gtggtcggcc acaggcggcg tggcgatcct
     7561 gtctccatcc gtctcgtcga ccgggaggcg agttcgatga ccctgataga gggggctgcg
     7621 gcttagggag gcagaagctg agcaccgtcg gtaggagctc cagggcccgg agcgactgac
     7681 ccctgccgag aactcagagg gtcgtcggaa gacggagagt gagcccgacg accaccccag
     7741 gcacgtcttt ggtcggcctg cggatcaagc tagcaagccg tcattaaggt gatttcgtcc
     7801 gcgtgtaaaa cctattgcgg gaaaatctct ccttctaaga aggaaatagg ggccatgttg
     7861 tccctgttac aaaaggaagg gttgctttag tctccctcag atttatattc tccggggtcc
     7921 tgggatccca tcactgcggc gctctcccag cgggcaatgg tacttggaaa atcgggagag
     7981 ttaaaaacct ggggattggt tttgggggca ttgaaggcgg ctcgagagta acaggactaa
     8041 cttgaatcga tgcgctaccg gactcagatc ttgccacc
//
