LOCUS       V024294                10354 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024294
VERSION     V024294
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..5050
                     /label="BACH1(NM_206866)"
                     /note="BACH1(NM_206866)"
                     /gene="BACH1"
     CDS             5057..5122
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5142..5641
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5662..6261
                     /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    6275..6863
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7070..7703
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7832..7848)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7856..7872
                     /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(7880..7910)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7925..7946
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8234..8822)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8993..9853)
                     /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(9854..9958)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10006..10140
                     /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 ccatgtctct gagtgagaac tcggtttttg cctatgaatc
     2881 ttctgtgcat agcaccaatg ttttactcag ccttaatgac cagcggaaga aagatgtgct
     2941 gtgcgatgtc accatctttg tggagggaca gcggttccgc gctcaccggt ccgtgctggc
     3001 ggcatgcagc agttacttcc actcaagaat cgtaggccag gctgatggag agctgaacat
     3061 tactcttcca gaagaggtga cagttaaagg atttgaacct ttaattcagt ttgcctacac
     3121 tgctaaactg attttaagta aagagaatgt ggatgaagtg tgcaaatgtg tggagttttt
     3181 aagtgtacat aatattgagg aatcctgctt tcagtttctg aaatttaagt ttttggactc
     3241 cactgcagac cagcaagaat gcccaagaaa aaaatgcttt tcatcacact gtcagaaaac
     3301 agaccttaaa ctttcacttt tggaccagag ggatctagaa actgatgaag tggaggaatt
     3361 tctggaaaat aaaaatgttc agactcctca gtgtaaactc cgcaggtatc aaggaaatgc
     3421 aaaagcctca cctcctctac aagacagtgc cagtcagaca tatgagtcca tgtgcttaga
     3481 gaaggatgct gctctggcct tgccttcttt atgccccaaa tacagaaaat tccaaaaagc
     3541 atttggaact gacagagtcc gtactgggga atctagtgtc aaagacattc atgcttctgt
     3601 tcagccaaat gaaaggtctg aaaatgaatg cctgggagga gtcccggagt gtagagattt
     3661 gcaggtgatg ttaaaatgtg acgaaagtaa attagcaatg gaacctgaag aaacgaagaa
     3721 agatcctgct tctcagtgcc caactgaaaa atcagaagtg actcctttcc cccacaattc
     3781 ttccatagac cctcatggac tttattcttt gtctctttta cacacatatg accaatatgg
     3841 tgacttgaat tttgctggta tgcaaaacac aacagtgtta acagaaaagc ctttgtcagg
     3901 tacagacgtc caagaaaaaa catttggtga aagtcaggat ttacctttga aatccgactt
     3961 gggcaccagg gaagatagta gtgttgcatc tagtgatagg agtagtgtgg agcgagaagt
     4021 ggcagaacac ctagcaaaag gcttctggag tgacatttgc agcacggaca ctccttgcca
     4081 aatgcagtta tcacctgctg tggccaaaga tggctcagaa cagatctcac agaaacggtc
     4141 tgagtgtccg tggttaggta tcaggattag tgagagccca gaaccaggtc aaaggacttt
     4201 cacaacatta agttctgtca actgcccttt tataagtact ctgagtactg aaggctgttc
     4261 aagcaatttg gaaattggaa acgatgatta tgtttcagaa ccccagcaag aaccttgccc
     4321 atatgcttgt gtcattagct tgggagacga ctctgagacg gacaccgaag gagacagtga
     4381 atcctgttca gccagagaac aagaatgtga ggtaaaactg ccattcaatg cacaacggat
     4441 aatttcactg tctcgaaatg attttcagtc cttgttgaaa atgcacaagc ttactccaga
     4501 acagctggat tgtatccatg atattcgaag aagaagtaaa aacagaattg ctgcacagcg
     4561 ctgtcgcaag agaaaacttg actgtataca gaatcttgaa tcagaaattg agaagctgca
     4621 aagtgaaaag gagagcttgt tgaaggaaag agatcacatt ttgtcaactc tgggtgagac
     4681 aaagcagaac ctaactggac tttgccagaa agtttgtaaa gaagcagctc tgagtcaaga
     4741 acaaatacag atactcgcca agtactcagc tgcagattgc ccactttcat ttttaatttc
     4801 tgaaaaagat aaaagtactc ctgatggtga actggcgtta ccatcaattt tcagtttatc
     4861 tgaccggcct ccagcagtgc tgcctccctg tgccagagga aacagtgagc ctggctacgc
     4921 gcgagggcag gagtcccagc agatgtccac agccacctct gagcaagctg ggcctgcgga
     4981 acagtgtcgt cagagtggtg ggatctcaga tttctgtcag cagatgactg ataaatgtac
     5041 tactgatgag ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga
     5101 ctacaaggat gacgatgaca agtagtctag ataattctac cgggtagggg aggcgctttt
     5161 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     5221 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     5281 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     5341 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     5401 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     5461 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     5521 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     5581 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     5641 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     5701 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     5761 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     5821 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     5881 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5941 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     6001 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     6061 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     6121 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     6181 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     6241 acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt acaaaatttg
     6301 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     6361 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     6421 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     6481 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     6541 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     6601 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6661 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     6721 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6781 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6841 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat
     6901 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     6961 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     7021 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta
     7081 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     7141 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga
     7201 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag
     7261 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg
     7321 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg
     7381 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt
     7441 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa
     7501 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     7561 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7621 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     7681 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     7741 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gttttaccgt
     7801 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7861 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7921 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7981 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     8041 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     8101 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     8161 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     8221 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     8281 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     8341 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     8401 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     8461 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8521 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8581 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8641 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8701 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8761 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8821 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8881 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8941 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     9001 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     9061 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     9121 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     9181 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     9241 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     9301 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     9361 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     9421 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     9481 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     9541 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     9601 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     9661 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9721 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9781 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9841 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9901 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9961 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca
    10021 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
    10081 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
    10141 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc
    10201 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca
    10261 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt
    10321 gtatttgtta aatatgtact acaaacttag tagt
//
