LOCUS       V033405                 9583 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033405
VERSION     V033405
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"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4437
                     /label="ORC3(NM_181837)"
                     /note="ORC3(NM_181837)"
                     /gene="ORC3"
     CDS             4444..4509
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4553..4764
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4777..5045
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     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"
     misc_feature    5678..6266
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6340..6573
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6645..6766
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6785..6920
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6958..6974)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6982..6998
                     /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(7006..7036)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7051..7072
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7360..7948)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8119..8979)
                     /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(8980..9084)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9558..9574
                     /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 gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggctacg tcctcgatgt ctaagggttg ctttgttttt
     2341 aagccaaact ccaaaaagag aaagatctct ctgccaatag aggactattt taacaaaggg
     2401 aaaaatgagc ctgaggacag taagcttcga ttcgaaactt atcagttgat atggcagcag
     2461 atgaaatctg aaaatgagcg actacaagag gaattaaata aaaacttgtt tgacaatctg
     2521 attgaatttc tgcaaaaatc acattctgga ttccagaaga attcaagaga cttgggcggt
     2581 caaataaaac tcagagaaat tccaactgct gctcttgttc ttggtgtgaa tgtcacagat
     2641 catgatttga cattcggaag tctaacagag gcccttcaga ataatgtcac accatatgta
     2701 gtctcattgc aagctaaaga ttgtccagat atgaaacatt ttttgcaaaa gttgatctca
     2761 cagttgatgg actgctgtgt agatataaaa tccaaagagg aggaaagtgt tcacgtcacc
     2821 caaagaaaga cacattattc aatggattca ctttccagtt ggtatatgac tgtcacacag
     2881 aagacggacc caaaaatgct aagcaaaaaa aggactactt ctagccaatg gcagtctcct
     2941 cctgttgtcg ttatcttgaa ggatatggaa agctttgcca caaaagtact acaagacttc
     3001 ataattatca gcagtcaaca tctccatgaa tttccactaa tactcatttt tggaatagcc
     3061 acatctccta ttatcatcca ccgattgctt cctcatgcag tatcatctct attgtgcata
     3121 gaactgttcc aatctttgtc ttgtaaggag cacctgacta cggtactcga taagctactt
     3181 cttacaactc agtttccctt taaaataaat gaaaaagtat tacaggttct gaccaacatc
     3241 tttttgtatc atgatttctc agttcaaaac tttataaaag gacttcagct ttctctatta
     3301 gagcatttct attcccagcc cttaagtgtc ctgtgctgta atcttccaga agccaaaaga
     3361 agaataaatt ttttatcaaa taatcaatgt gaaaacatcc gacgtctacc atcttttagg
     3421 aggtacgtgg aaaagcaagc ttcagaaaag caagttgcgc tcttgaccaa tgagagatat
     3481 ttgaaggagg aaacacaatt attactagaa aacctgcatg tttatcatat gaattacttc
     3541 ctggttttga gatgtcttca taagttcacc tcttctcttc ccaagtatcc actaggtcga
     3601 cagatcagag agttgtactg tacatgttta gaaaagaaca tatgggattc agaggagtat
     3661 gcatcagtct tgcagctgct gaggatgttg gcaaaggatg aactgatgac catacttgag
     3721 aaatgtttca aggtttttaa gtcttattgt gaaaaccacc ttggcagcac agctaagaga
     3781 atagaggagt tcctggccca gtttcagagc ctcgatgcag aaaccaaaga ggaagaagat
     3841 gcttctgggt cacagccaaa ggggcttcag aagacagacc tctatcatct tcagaagtcc
     3901 ttattggaaa tgaaggagtt aagaagaagt aagaagcaaa ccaaatttga agtactcaga
     3961 gaaaatgttg tgaacttcat tgactgtcta gtgagagaat accttctgcc tcctgagaca
     4021 cagcctctcc atgaggtggt gtacttcagt gctgcccatg cccttcgtga gcatttaaat
     4081 gctgctccgc gaattgccct ccatactgca ctcaacaatc cttactatta tctcaagaat
     4141 gaagcactga aaagcgaaga aggctgcatt ccgaatatcg ccccagacat ctgcatagca
     4201 tacaaactgc acctagagtg tagcaggctc atcaacctcg tggactggtc agaggctttt
     4261 gcaacagttg tgacagctgc tgaaaaaatg gatgcaaatt ctgcaacctc agaagaaatg
     4321 aatgaaatta tccatgctcg gtttattaga gctgtttctg aactagaact tttaggattt
     4381 ataaaaccta ccaaacagaa gactgaccat gtggcaagac taacatgggg aggctgcgga
     4441 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     4501 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     4561 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     4621 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     4681 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     4741 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     4801 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     4861 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     4921 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     4981 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     5041 agatccaagc tgtgaccggc gcctacgcta gatgaccgag tacaagccca cggtgcgcct
     5101 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     5161 ccccgccacg cgccacaccg tcgacccgga 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 agtcgacaat caacctctgg attacaaaat
     5701 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5761 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5821 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5881 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5941 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6001 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6061 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     6121 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6181 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6241 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     6301 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6361 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     6421 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6481 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6541 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6601 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6661 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6721 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6781 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6841 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6901 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6961 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     7021 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7081 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7141 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7201 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7261 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7321 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7681 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7741 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7801 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7861 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7921 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7981 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     8041 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     8101 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8161 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8221 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8281 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8341 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8401 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8461 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8521 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8581 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8641 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8701 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8761 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8821 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8881 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8941 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     9001 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9061 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     9121 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     9181 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     9241 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     9301 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9361 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     9421 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9481 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9541 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
