LOCUS       V028532                 9339 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028532
VERSION     V028532
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..4008
                     /label="HTR2B(NM_000867)"
                     /note="HTR2B(NM_000867)"
                     /gene="HTR2B"
     misc_feature    4054..4642
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     promoter        4673..5172
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5189..5788
                     /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             5860..6093
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6165..6286
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6326..6461
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6482..6500)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6510..6526)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      6668..7123
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7149..7253
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7254..8114
                     /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      8285..8873
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9161..9182
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        9197..9227
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9235..9251
                     /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     9259..9275
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        9296..9314
                     /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 ccaccatggc tctctcttac
     2581 agagtgtctg aacttcaaag cacaattcct gagcacattt tgcagagcac ctttgttcac
     2641 gttatctctt ctaactggtc tggattacag acagaatcaa taccagagga aatgaaacag
     2701 attgttgagg aacagggaaa taaactgcac tgggcagctc ttctgatact catggtgata
     2761 atacccacaa ttggtggaaa tacccttgtt attctggctg tttcactgga gaagaagctg
     2821 cagtatgcta ctaattactt tctaatgtcc ttggcggtgg ctgatttgct ggttggattg
     2881 tttgtgatgc caattgccct cttgacaata atgtttgagg ctatgtggcc cctcccactt
     2941 gttctatgtc ctgcctggtt atttcttgac gttctctttt caaccgcatc catcatgcat
     3001 ctctgtgcca tttcagtgga tcgttacata gccatcaaaa agccaatcca ggccaatcaa
     3061 tataactcac gggctacagc attcatcaag attacagtgg tgtggttaat ttcaataggc
     3121 attgccattc cagtccctat taaagggata gagactgatg tggacaaccc aaacaatatc
     3181 acttgtgtgc tgacaaagga acgttttggc gatttcatgc tctttggctc actggctgcc
     3241 ttcttcacac ctcttgcaat tatgattgtc acctactttc tcactatcca tgctttacag
     3301 aagaaggctt acttagtcaa aaacaagcca cctcaacgcc taacatggtt gactgtgtct
     3361 acagttttcc aaagggatga aacaccttgc tcgtcaccgg aaaaggtggc aatgctggat
     3421 ggttctcgaa aggacaaggc tctgcccaac tcaggtgatg aaacacttat gcgaagaaca
     3481 tccacaattg ggaaaaagtc agtgcagacc atttccaacg aacagagagc ctcaaaggtc
     3541 ctagggattg tgtttttcct ctttttgctt atgtggtgtc ccttctttat tacaaatata
     3601 actttagttt tatgtgattc ctgtaaccaa actactctcc aaatgctcct ggagatattt
     3661 gtgtggatag gctatgtttc ctcaggagtg aatcctttgg tctacaccct cttcaataag
     3721 acatttcggg atgcatttgg ccgatatatc acctgcaatt accgggccac aaagtcagta
     3781 aaaactctca gaaaacgctc cagtaagatc tacttccgga atccaatggc agagaactct
     3841 aagtttttca agaaacatgg aattcgaaat gggattaacc ctgccatgta ccagagtcca
     3901 atgaggctcc gaagttcaac cattcagtct tcatcaatca ttctactaga tacgcttctc
     3961 ctcactgaaa atgaaggtga caaaactgaa gagcaagtta gttatgtata gctcgagtct
     4021 agatgtacaa agtggtgata atcgaattcc gataatcaac ctctggatta caaaatttgt
     4081 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4141 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4201 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4261 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4321 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4381 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4441 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     4501 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4561 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4621 tccctttggg ccgcctcccc gcatcgggaa ttcccgcggt tcgaattcta ccgggtaggg
     4681 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
     4741 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
     4801 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
     4861 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
     4921 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     4981 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     5041 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     5101 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     5161 ccgggccttt cgacctcacg tggccaccat gaccgagtac aagcccacgg tgcgcctcgc
     5221 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5281 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5341 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5401 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5461 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5521 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5581 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5641 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5701 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5761 gtgcatgacc cgcaagcccg gtgcctgagg tacctttaag accaatgact tacaaggcag
     5821 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     5881 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     5941 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6001 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6061 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6121 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6181 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6241 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6301 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     6481 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     6541 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     6601 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     6661 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6721 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6781 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6841 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6901 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6961 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     7021 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     7081 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     7141 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     7201 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     7261 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     7321 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     7381 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     7441 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7501 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7561 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7621 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7681 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7741 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7801 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7861 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7921 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7981 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     8041 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     8101 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     8161 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8221 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     8281 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     8341 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     8401 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     8461 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     8521 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8581 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8641 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8701 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8761 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8821 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8881 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8941 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     9001 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     9061 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     9121 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     9181 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     9241 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     9301 accctcacta aagggaacaa aagctggagc tgcaagctt
//
