LOCUS       V028674                 9222 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028674
VERSION     V028674
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..3891
                     /label="SDC3(NM_014654)"
                     /note="SDC3(NM_014654)"
                     /gene="SDC3"
     misc_feature    3937..4525
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     promoter        4556..5055
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5072..5671
                     /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             5743..5976
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6048..6169
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6209..6344
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6365..6383)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6393..6409)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      6551..7006
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7032..7136
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7137..7997
                     /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      8168..8756
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9044..9065
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        9080..9110
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9118..9134
                     /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     9142..9158
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        9179..9197
                     /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 ccaccatgaa gccggggccg
     2581 ccgcaccgtg ccggggccgc ccacggggcc ggcgccgggg ccggggccgc ggccgggccc
     2641 ggggcccgcg ggctgctcct gccaccgctg ctgctgctgc tgctggcggg gcgcgccgcg
     2701 ggggcccagc gctggcgcag tgagaacttc gagagacccg tggacctgga gggctctggg
     2761 gatgatgact cctttcccga tgatgaactg gatgacctct actcggggtc gggctcgggc
     2821 tacttcgagc aggagtcggg cattgagaca gccatgcgct tcagcccaga tgtagccctg
     2881 gcggtgtcca ccacacctgc ggtgctgccc accacgaaca tccagcctgt gggcacacca
     2941 tttgaagagc tcccctctga gcgccccacc ctggagccag ccaccagccc cctggtggtg
     3001 acagaagtcc cggaagagcc cagccagaga gccaccaccg tctccactac catggctacc
     3061 actgctgcca caagcacagg ggacccgact gtggccacag tgcctgccac agtggccacc
     3121 gccaccccca gcacccctgc agcaccccct tttacggcca ccactgctgt tataaggacc
     3181 actggcgtac ggaggcttct gcctctccca ctgaccacag tggctacggc acgggccact
     3241 acccccgagg cgccctcccc gcccaccacg gcggctgtct tggacaccga ggccccaaca
     3301 cccaggctgg tcagcacagc tacctcccgg ccaagagccc ttcccaggcc ggccaccacc
     3361 caggagcctg acatccctga gaggagcacc ctgcccctgg ggaccactgc ccctggaccc
     3421 acagaggtgg ctcagacccc aactccagag accttcctga ccacaatccg ggatgagcca
     3481 gaggttccgg tgagtggggg gcccagtgga gacttcgagc tgccagaaga agagaccaca
     3541 caaccagaca cagccaatga ggtggtagct gtgggagggg ctgcggccaa ggcatcatct
     3601 ccacctggga cactgcccaa gggtgcccgc ccgggccctg gcctcctgga caatgccatc
     3661 gactcgggca gctcagctgc tcagctgcct cagaagagta tcctggagcg gaaggaggtg
     3721 ctcgtagctg tgattgtggg cggggtggtg ggcgccctct ttgctgcctt cttggtcaca
     3781 ctgctcatct atcgtatgaa gaaaaaggat gagggcagct acacgctgga ggaacccaag
     3841 caggcgagcg tcacatacca gaagcctgac aagcaggagg agttctatgc ctagctcgag
     3901 tctagatgta caaagtggtg ataatcgaat tccgataatc aacctctgga ttacaaaatt
     3961 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4021 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     4081 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     4141 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4201 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     4261 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     4321 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     4381 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     4441 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     4501 atctcccttt gggccgcctc cccgcatcgg gaattcccgc ggttcgaatt ctaccgggta
     4561 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     4621 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     4681 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     4741 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     4801 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     4861 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     4921 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     4981 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     5041 tctccgggcc tttcgacctc acgtggccac catgaccgag tacaagccca cggtgcgcct
     5101 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     5161 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     5221 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     5281 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     5341 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     5401 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     5461 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     5521 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     5581 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     5641 ctggtgcatg acccgcaagc ccggtgcctg aggtaccttt aagaccaatg acttacaagg
     5701 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5761 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5821 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5881 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5941 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6001 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6061 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6121 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6181 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6241 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6301 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     6361 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     6421 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     6481 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     6541 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6601 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6661 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6721 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6781 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6841 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     6901 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6961 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     7021 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     7081 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     7141 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     7201 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     7261 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     7321 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     7381 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     7441 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     7501 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7561 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7621 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7681 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7741 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7801 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7861 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7921 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7981 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     8041 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     8101 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8161 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     8221 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8281 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     8341 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     8401 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8461 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8521 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8581 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8641 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8701 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8761 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8821 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8881 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8941 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     9001 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     9061 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     9121 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     9181 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc tt
//
