LOCUS       V035528                 9683 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035528
VERSION     V035528
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        1933..2236
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2237..2440
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2497..5034
                     /label="EXO1(NM_006027)"
                     /note="EXO1(NM_006027)"
                     /gene="EXO1"
     CDS             5071..5772
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric blue fluorescent protein"
                     /transl_table=1
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN*"
                     /label="TagBFP"
     misc_feature    5779..6366
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6440..6673
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6745..6866
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6885..7020
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7058..7074)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7082..7098
                     /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"
     promoter        complement(7106..7136)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7151..7172
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7460..8048)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8219..9079)
                     /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"
     promoter        complement(9080..9184)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9658..9674
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga 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 ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1981 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     2041 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2101 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2161 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2221 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2281 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2341 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     2401 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     2461 ccgctagcgc taccggactc agatctcgag gccaccatgg ggatacaggg attgctacaa
     2521 tttatcaaag aagcttcaga acccatccat gtgaggaagt ataaagggca ggtagtagct
     2581 gtggatacat attgctggct tcacaaagga gctattgctt gtgctgaaaa actagccaaa
     2641 ggtgaaccta ctgataggta tgtaggattt tgtatgaaat ttgtaaatat gttactatct
     2701 catgggatca agcctattct cgtatttgat ggatgtactt taccttctaa aaaggaagta
     2761 gagagatcta gaagagaaag acgacaagcc aatcttctta agggaaagca acttcttcgt
     2821 gaggggaaag tctcggaagc tcgagagtgt ttcacccggt ctatcaatat cacacatgcc
     2881 atggcccaca aagtaattaa agctgcccgg tctcaggggg tagattgcct cgtggctccc
     2941 tatgaagctg atgcgcagtt ggcctatctt aacaaagcgg gaattgtgca agccataatt
     3001 acagaggact cggatctcct agcttttggc tgtaaaaagg taattttaaa gatggaccag
     3061 tttggaaatg gacttgaaat tgatcaagct cggctaggaa tgtgcagaca gcttggggat
     3121 gtattcacgg aagagaagtt tcgttacatg tgtattcttt caggttgtga ctacctgtca
     3181 tcactgcgtg ggattggatt agcaaaggca tgcaaagtcc taagactagc caataatcca
     3241 gatatagtaa aggttatcaa gaaaattgga cattatctca agatgaatat cacggtacca
     3301 gaggattaca tcaacgggtt tattcgggcc aacaatacct tcctctatca gctagttttt
     3361 gatcccatca aaaggaaact tattcctctg aacgcctatg aagatgatgt tgatcctgaa
     3421 acactaagct acgctgggca atatgttgat gattccatag ctcttcaaat agcacttgga
     3481 aataaagata taaatacttt tgaacagatc gatgactaca atccagacac tgctatgcct
     3541 gcccattcaa gaagtcatag ttgggatgac aaaacatgtc aaaagtcagc taatgttagc
     3601 agcatttggc ataggaatta ctctcccaga ccagagtcgg gtactgtttc agatgcccca
     3661 caattgaagg aaaatccaag tactgtggga gtggaacgag tgattagtac taaagggtta
     3721 aatctcccaa ggaaatcatc cattgtgaaa agaccaagaa gtgcagagct gtcagaagat
     3781 gacctgttga gtcagtattc tctttcattt acgaagaaga ccaagaaaaa tagctctgaa
     3841 ggcaataaat cattgagctt ttctgaagtg tttgtgcctg acctggtaaa tggacctact
     3901 aacaaaaaga gtgtaagcac tccacctagg acgagaaata aatttgcaac atttttacaa
     3961 aggaaaaatg aagaaagtgg tgcagttgtg gttccaggga ccagaagcag gtttttttgc
     4021 agttcagatt ctactgactg tgtatcaaac aaagtgagca tccagcctct ggatgaaact
     4081 gctgtcacag ataaagagaa caatctgcat gaatcagagt atggagacca agaaggcaag
     4141 agactggttg acacagatgt agcacgtaat tcaagtgatg acattccgaa taatcatatt
     4201 ccaggtgatc atattccaga caaggcaaca gtgtttacag atgaagagtc ctactctttt
     4261 gagagcagca aatttacaag gaccatttca ccacccactt tgggaacact aagaagttgt
     4321 tttagttggt ctggaggtct tggagatttt tcaagaacgc cgagcccctc tccaagcaca
     4381 gcattgcagc agttccgaag aaagagcgat tcccccacct ctttgcctga gaataatatg
     4441 tctgatgtgt cgcagttaaa gagcgaggag tccagtgacg atgagtctca tcccttacga
     4501 gaagaggcat gttcttcaca gtcccaggaa agtggagaat tctcactgca gagttcaaat
     4561 gcatcaaagc tttctcagtg ctctagtaag gactctgatt cagaggaatc tgattgcaat
     4621 attaagttac ttgacagtca aagtgaccag acctccaagc tacgtttatc tcatttctca
     4681 aaaaaagaca cacctctaag gaacaaggtt cctgggctat ataagtccag ttctgcagac
     4741 tctctttcta caaccaagat caaacctcta ggacctgcca gagccagtgg gctgagcaag
     4801 aagccggcaa gcatccagaa gagaaagcat cataatgccg agaacaagcc ggggttacag
     4861 atcaaactca atgagctctg gaaaaacttt ggatttaaaa aagattctga aaagcttcct
     4921 ccttgtaaga aacccctgtc cccagtcaga gataacatcc aactaactcc agaagcggaa
     4981 gaggatatat ttaacaaacc tgaatgtggc cgtgttcaaa gagcaatatt ccaggaattc
     5041 ggcggtggag gaagcggcgg aggtggatct atgagcgagc tgattaagga gaacatgcac
     5101 atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc
     5161 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
     5221 ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc
     5281 aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg
     5341 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
     5401 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc
     5461 cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct
     5521 gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg
     5581 atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct
     5641 ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac
     5701 gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac
     5761 aagcttaatt aaggatccaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5821 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5881 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5941 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6001 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6061 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6121 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     6181 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6241 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6301 cggcctcttc cgcgtctccg ccttcgccct cagacgagtc ggatctccct ttggccgcct
     6361 ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6421 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt
     6481 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6541 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6601 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6661 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6721 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     6781 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6841 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     6901 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6961 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7021 tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa
     7081 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7141 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7201 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7261 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7321 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7381 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7441 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7501 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7561 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7621 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7681 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7741 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7801 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7861 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7921 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7981 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8041 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8101 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8161 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8221 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8281 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8341 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8401 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8461 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8521 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8581 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8641 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8701 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8761 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8821 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8881 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8941 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9001 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9061 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9121 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9181 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     9241 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     9301 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     9361 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     9421 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     9481 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     9541 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     9601 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     9661 aaacgacggc cagtgccaag ctg
//
