LOCUS       V028346                 9417 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028346
VERSION     V028346
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        3..229
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             230..410
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    457..582
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1075..1308
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1803..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     protein_bind    1938..1958
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of attB"
                     /note="core recombination site for the Gateway® BP
                     reaction"
                     /label="attB4"
     enhancer        2019..2322
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2323..2526
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2566..4086
                     /label="TGFBR1(NM_001306210)"
                     /note="TGFBR1(NM_001306210)"
                     /gene="TGFBR1"
     misc_feature    4132..4720
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     promoter        4751..5250
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5267..5866
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     LTR             5938..6171
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6243..6364
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6404..6539
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6560..6578)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6588..6604)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      6746..7201
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7227..7331
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7332..8192
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      8363..8951
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9239..9260
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        9275..9305
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9313..9329
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     primer_bind     9337..9353
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        9374..9392
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
     1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1921 actagtgatt atcggatcaa ctttgtatag aaaagttgta gttattaata gtaatcaatt
     1981 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2041 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2101 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2161 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2221 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2281 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2341 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2401 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2461 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2521 agagctggtt tagtgaaccg tcagatccaa gttggatccg ccaccatgga ggcggcggtc
     2581 gctgctccgc gtccccggct gctcctcctc gtgctggcgg cggcggcggc ggcggcggcg
     2641 gcgctgctcc cgggggcgac ggcgttacag tgtttctgcc acctctgtac aaaagacaat
     2701 tttacttgtg tgacagatgg gctctgcttt gtctctgtca cagagaccac agacaaagtt
     2761 atacacaaca gcatgtgtat agctgaaatt gacttaattc ctcgagatag gccgtttgta
     2821 tgtgcaccct cttcaaaaac tgggtctgtg actacaacat attgctgcaa tcaggaccat
     2881 tgcaataaaa tagaacttcc aactactggc cctttttcag taaagtcatc acctggcctt
     2941 ggtcctgtgg aactggcagc tgtcattgct ggaccagtgt gcttcgtctg catctcactc
     3001 atgttgatgg tctatatctg ccacaaccgc actgtcattc accatcgagt gccaaatgaa
     3061 gaggaccctt cattagatcg cccttttatt tcagagggta ctacgttgaa agacttaatt
     3121 tatgatatga caacgtcagg ttctggctca ggtttaccat tgcttgttca gagaacaatt
     3181 gcgagaacta ttgtgttaca agaaagcatt ggcaaaggtc gatttggaga agtttggaga
     3241 ggaaagtggc ggggagaaga agttgctgtt aagatattct cctctagaga agaacgttcg
     3301 tggttccgtg aggcagagat ttatcaaact gtaatgttac gtcatgaaaa catcctggga
     3361 tttatagcag cagacaataa agacaatggt acttggactc agctctggtt ggtgtcagat
     3421 tatcatgagc atggatccct ttttgattac ttaaacagat acacagttac tgtggaagga
     3481 atgataaaac ttgctctgtc cacggcgagc ggtcttgccc atcttcacat ggagattgtt
     3541 ggtacccaag gaaagccagc cattgctcat agagatttga aatcaaagaa tatcttggta
     3601 aagaagaatg gaacttgctg tattgcagac ttaggactgg cagtaagaca tgattcagcc
     3661 acagatacca ttgatattgc tccaaaccac agagtgggaa caaaaaggta catggcccct
     3721 gaagttctcg atgattccat aaatatgaaa cattttgaat ccttcaaacg tgctgacatc
     3781 tatgcaatgg gcttagtatt ctgggaaatt gctcgacgat gttccattgg tggaattcat
     3841 gaagattacc aactgcctta ttatgatctt gtaccttctg acccatcagt tgaagaaatg
     3901 agaaaagttg tttgtgaaca gaagttaagg ccaaatatcc caaacagatg gcagagctgt
     3961 gaagccttga gagtaatggc taaaattatg agagaatgtt ggtatgccaa tggagcagct
     4021 aggcttacag cattgcggat taagaaaaca ttatcgcaac tcagtcaaca ggaaggcatc
     4081 aaaatgtaac tcgagtctag atgtacaaag tggtgataat cgaattccga taatcaacct
     4141 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4201 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4261 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     4321 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     4381 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     4441 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     4501 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     4561 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     4621 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4681 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgggaatt cccgcggttc
     4741 gaattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca
     4801 gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc
     4861 accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct
     4921 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg
     4981 gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg
     5041 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg
     5101 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc
     5161 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
     5221 ttctcctctt cctcatctcc gggcctttcg acctcacgtg gccaccatga ccgagtacaa
     5281 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     5341 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     5401 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     5461 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     5521 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     5581 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5641 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5701 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5761 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5821 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaggta cctttaagac
     5881 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5941 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     6001 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6061 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6121 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6181 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6241 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6301 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6361 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     6421 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6481 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6541 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     6601 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     6661 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     6721 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     6781 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6841 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6901 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6961 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     7021 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     7081 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     7141 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     7201 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7261 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7321 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7381 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7441 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7501 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7561 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7621 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7681 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7741 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7801 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7861 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7921 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7981 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     8041 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     8101 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     8161 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     8221 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     8281 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     8341 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     8401 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     8461 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     8521 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     8581 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     8641 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     8701 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8761 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8821 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8881 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8941 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     9001 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     9061 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     9121 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     9181 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     9241 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     9301 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     9361 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagctt
//
