LOCUS       V044071                 9424 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044071
VERSION     V044071
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..4120
                     /label="EGLN1(NM_022051)"
                     /note="EGLN1(NM_022051)"
                     /gene="EGLN1"
     CDS             4127..4192
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4212..4711
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4732..5331
                     /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    5345..5933
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6140..6773
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6902..6918)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6926..6942
                     /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(6950..6980)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6995..7016
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7304..7892)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8063..8923)
                     /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(8924..9028)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9076..9210
                     /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 ccatggccaa tgacagcggc gggcccggcg ggccgagccc
     2881 gagcgagcga gaccggcagt actgcgagct gtgcgggaag atggagaacc tgctgcgctg
     2941 cagccgctgc cgcagctcct tctactgctg caaggagcac cagcgtcagg actggaagaa
     3001 gcacaagctc gtgtgccagg gcagcgaggg cgccctcggc cacggagtgg gcccacacca
     3061 gcattccggc cccgcgccgc cggctgcagt gccgccgccc agggccgggg cccgggagcc
     3121 caggaaggca gcggcgcgcc gggacaacgc ctccggggac gcggccaagg gaaaagtaaa
     3181 ggccaagccc ccggccgacc cagcggcggc cgcgtcgccg tgtcgtgcgg ccgccggcgg
     3241 ccagggctcg gcggtggctg ccgaagccga gcccggcaag gaggagccgc cggcccgctc
     3301 atcgctgttc caggagaagg cgaacctgta ccccccaagc aacacgcccg gggatgcgct
     3361 gagccccggc ggcggcctgc ggcccaacgg gcagacgaag cccctgccgg cgctgaagct
     3421 ggcgctcgag tacatcgtgc cgtgcatgaa caagcacggc atctgtgtgg tggacgactt
     3481 cctcggcaag gagaccggac agcagatcgg cgacgaggtg cgcgccctgc acgacaccgg
     3541 gaagttcacg gacgggcagc tggtcagcca gaagagtgac tcgtccaagg acatccgagg
     3601 cgataagatc acctggatcg agggcaagga gcccggctgc gaaaccattg ggctgctcat
     3661 gagcagcatg gacgacctga tacgccactg taacgggaag ctgggcagct acaaaatcaa
     3721 tggccggacg aaagccatgg ttgcttgtta tccgggcaat ggaacgggtt atgtacgtca
     3781 tgttgataat ccaaatggag atggaagatg tgtgacatgt atatattatc ttaataaaga
     3841 ctgggatgcc aaggtaagtg gaggtatact tcgaattttt ccagaaggca aagcccagtt
     3901 tgctgacatt gaacccaaat ttgatagact gctgtttttc tggtctgacc gtcgcaaccc
     3961 tcatgaagta caaccagcat atgctacaag gtacgcaata actgtttggt attttgatgc
     4021 agatgagaga gcacgagcta aagtaaaata tctaacaggt gaaaaaggtg tgagggttga
     4081 actcaataaa ccttcagatt cggtcggtaa agacgtcttc ggatccgact acaaagacca
     4141 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtctag
     4201 ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc
     4261 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc
     4321 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc
     4381 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat
     4441 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg
     4501 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag
     4561 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc
     4621 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct
     4681 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag
     4741 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     4801 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     4861 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4921 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4981 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5041 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     5101 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     5161 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5221 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5281 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg
     5341 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5401 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5461 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5521 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5581 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5641 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5701 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     5761 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5821 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5881 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     5941 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct
     6001 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc
     6061 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6121 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt
     6181 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg
     6241 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca
     6301 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg
     6361 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca
     6421 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct
     6481 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg
     6541 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct
     6601 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     6661 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     6721 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     6781 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     6841 gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg gcgtaatcat
     6901 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6961 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     7021 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     7081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7741 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7921 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7981 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     8041 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     8101 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8161 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8221 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8281 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8341 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8401 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8461 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8521 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8581 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8641 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8701 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8761 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8821 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8881 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8941 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     9001 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     9061 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     9121 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     9181 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
     9241 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
     9301 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
     9361 actacaaact tagtagtttt taaagaaatt gtatttgtta aatatgtact acaaacttag
     9421 tagt
//
