LOCUS       V033429                 9610 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033429
VERSION     V033429
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2843..4306
                     /label="PDIA6(NM_001282705)"
                     /note="PDIA6(NM_001282705)"
                     /gene="PDIA6"
     CDS             4313..4378
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4398..4897
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4918..5517
                     /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    5531..6119
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6326..6959
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7088..7104)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7112..7128
                     /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(7136..7166)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7181..7202
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7490..8078)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8249..9109)
                     /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(9110..9214)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9262..9396
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ctcaagcttc gaattcgcca ccatgaggat aatcacagcc cctgcctcca aagtgtcccg
     2881 aggatcaaat gagttgatga tacttgcacg gcgctcggat cgtggctcac caacttctcc
     2941 tgcccactcc ctatcaagga aaagtcccat aatgtatcca tcaaccacca tggctaatgc
     3001 acccggtctg gtgagctgta ccttctttct ggcagtgaat ggtctgtatt cctctagtga
     3061 tgatgtgatc gaattaactc catcgaattt caaccgagaa gttattcaga gtgatagttt
     3121 gtggcttgta gaattctatg ctccatggtg tggtcactgt caaagattaa caccagaatg
     3181 gaagaaagca gcaactgcat taaaagatgt tgtcaaagtt ggtgcagttg atgcagataa
     3241 gcatcattcc ctaggaggtc agtatggtgt tcagggattt cctaccatta agatttttgg
     3301 atccaacaaa aacagaccag aagattacca aggtggcaga actggtgaag ccattgtaga
     3361 tgctgcgctg agtgctctgc gccagctcgt gaaggatcgc ctcgggggac ggagcggagg
     3421 atacagttct ggaaaacaag gcagaagtga tagttcaagt aagaaggatg tgattgagct
     3481 gacagacgac agctttgata agaatgttct ggacagtgaa gatgtttgga tggttgagtt
     3541 ctatgctcct tggtgtggac actgcaaaaa cctagagcca gagtgggctg ccgcagcttc
     3601 agaagtaaaa gagcagacga aaggaaaagt gaaactggca gctgtggatg ctacagtcaa
     3661 tcaggttctg gcctcccgat acgggattag aggatttcct acaatcaaga tatttcagaa
     3721 aggcgagtct cctgtggatt atgacggtgg gcggacaaga tccgacatcg tgtcccgggc
     3781 ccttgatttg ttttctgata acgccccacc tcctgagctg cttgagatta tcaacgagga
     3841 cattgccaag aggacgtgtg aggagcacca gctctgtgtt gtggctgtgc tgccccatat
     3901 ccttgatact ggagctgcag gcagaaattc ttatctggaa gttcttctga agttggcaga
     3961 caaatacaaa aagaaaatgt gggggtggct gtggacagaa gctggagccc agtctgaact
     4021 tgagaccgcg ttggggattg gagggtttgg gtaccccgcc atggccgcca tcaatgcacg
     4081 caagatgaaa tttgctctgc taaaaggctc cttcagtgag caaggcatca acgagtttct
     4141 cagggagctc tcttttgggc gtggctccac ggcacctgta ggaggcgggg ctttccctac
     4201 catcgttgag agagagcctt gggacggcag ggatggcgag cttcccgtgg aggatgacat
     4261 tgacctcagt gatgtggagc ttgatgactt agggaaagat gagttgggat ccgactacaa
     4321 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     4381 gtctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
     4441 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
     4501 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
     4561 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
     4621 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
     4681 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
     4741 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
     4801 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
     4861 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg
     4921 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     4981 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     5041 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     5101 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     5161 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     5221 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     5281 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     5341 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     5401 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5461 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg
     5521 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5581 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5641 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5701 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5761 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5821 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5881 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5941 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6001 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     6061 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     6121 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca
     6181 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     6241 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     6301 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat
     6361 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca
     6421 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt
     6481 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg
     6541 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc
     6601 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat
     6661 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg
     6721 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg
     6781 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     6841 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     6901 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     6961 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     7021 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt
     7081 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     7141 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7201 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7261 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7321 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7381 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7441 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7501 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7561 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7621 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7681 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7741 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7801 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7861 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7921 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7981 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8041 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8101 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8161 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8221 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8281 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8341 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8401 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8461 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8521 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8581 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8641 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8701 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8761 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8821 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8881 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8941 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     9001 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9061 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9121 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9181 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9241 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc
     9301 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     9361 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
     9421 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
     9481 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
     9541 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa
     9601 acttagtagt
//
