LOCUS       V045429                 9121 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045429
VERSION     V045429
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"
     regulatory      2809..2818
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2818..2847
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2848..3792
                     /label="Ddrgk1(NM_029832)"
                     /note="Ddrgk1(NM_029832)"
                     /gene="Ddrgk1"
     misc_feature    3824..4410
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             4430..5029
                     /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    5043..5631
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5838..6471
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6599..6615)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6623..6639
                     /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(6647..6677)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6692..6713
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7001..7589)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7760..8620)
                     /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(8621..8725)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8773..8907
                     /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 tctatttccg gtgaattcgc caccatggag
     2821 cagaagctga tctcagagga ggacctgatg gtggggccct gggtgtatct ggtggcggca
     2881 gttttgctca tcggcctgat cctcttcctg actcgcagcc ggggtcgggc ggcagcagct
     2941 gacggagaac cactgcacaa tgaggaagag agggcaggag caggccaggt aggccgctct
     3001 ttgccccagg agtctgaaga acagagaact ggaagcagac cccggcgtcg gagggacttg
     3061 ggcagccgtc tacaggccca gcgtcgagcc cagcgagtgg cctgggaaga cggggatgag
     3121 aatgtgggtc aaactgttat tccagcccag gaggaagaag gcattgagaa gccagcagaa
     3181 gttcacccaa cagggaaaat tggagccaag aaactacgga agctagagga aaaacaggct
     3241 cgaaaggctc agcgagaggc agaggaggct gaacgtgaag aacggaaacg cctagagtcc
     3301 caacgtgagg ccgaatggaa gaaggaagag gaacggcttc gcctgaagga agaacagaag
     3361 gaggaggaag agaggaaggc tcaggaggag caggcccggc gggagcacga ggagtacctg
     3421 aaactgaagg aggccttcgt ggtagaagaa gaaggtgtta gcgaaaccat gactgaggag
     3481 cagtctcaca gcttcctgac agaattcatc aattacatca agaagtccaa ggttgtgctt
     3541 ttggaagatc tggctttcca gatgggccta aggactcagg acgccataaa ccgcatccag
     3601 gacctgctga cggaggggac tctaacaggt gtgattgacg accggggcaa gtttatctac
     3661 ataaccccag aggaactggc tgccgtggcc aatttcatcc gacagcgggg ccgggtgtcc
     3721 atcacagagc ttgcccaggc cagcaactcc ctcatctcct ggggccagga cctccctgcc
     3781 caggcttcag cctgaactag ttctagagcg gccgcggatc ccgcccctct ccctcccccc
     3841 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     3901 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     3961 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     4021 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     4081 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     4141 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     4201 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     4261 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     4321 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     4381 gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca tgaccgagta
     4441 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
     4501 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
     4561 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4621 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     4681 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     4741 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     4801 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     4861 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     4921 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     4981 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctaga cgcgtctgga
     5041 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5101 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5161 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5221 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5281 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5341 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5401 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     5461 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5521 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5581 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta
     5641 attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
     5701 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
     5761 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     5821 aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     5881 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
     5941 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     6001 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     6061 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     6121 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
     6181 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
     6241 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
     6301 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6361 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6421 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6481 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6541 ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg taatcatggt
     6601 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6661 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6721 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6841 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6901 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6961 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7021 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7081 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7141 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7201 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7261 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7321 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7381 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7441 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7501 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7561 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7621 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7681 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7741 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7801 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7861 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7921 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7981 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8041 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8101 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8161 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8221 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8281 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8341 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8401 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8461 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8521 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8581 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8641 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8701 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8761 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     8821 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     8881 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     8941 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     9001 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     9061 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     9121 t
//
