LOCUS       V044586                 9859 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044586
VERSION     V044586
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..4534
                     /label="KRT6B(NM_005555)"
                     /note="KRT6B(NM_005555)"
                     /gene="KRT6B"
     CDS             4541..4567
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4571..4597
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4601..4627
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4647..5146
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5167..5766
                     /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    5780..6368
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6575..7208
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7337..7353)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7361..7377
                     /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(7385..7415)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7430..7451
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7739..8327)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8498..9358)
                     /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(9359..9463)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9511..9645
                     /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 ccatggccag cacatccacc accatcagga gccacagcag
     2881 cagccgccgg ggtttcagtg ccaactcagc caggctccct ggggtcagcc gctctggctt
     2941 cagcagcatc tccgtgtccc gctccagggg cagtggtggc ctgggtggcg catgtggagg
     3001 agctggcttt ggcagccgca gtctgtatgg cctggggggc tccaagagga tctccattgg
     3061 agggggcagc tgtgccatca gtggcggcta tggcagcaga gccggaggca gctatggctt
     3121 tggtggcgcc gggagtggat ttggtttcgg tggtggagcc ggcattggct ttggtctggg
     3181 tggtggagcc ggccttgctg gtggctttgg gggccctggc ttccctgtgt gcccccctgg
     3241 aggcatccaa gaggtcactg tcaaccagag tctcctgact cccctcaacc tgcaaattga
     3301 ccccgccatc cagcgggtgc gggccgagga gcgtgagcag atcaagaccc tcaacaacaa
     3361 gtttgcctcc ttcatcgaca aggtgcggtt cctagagcag cagaacaagg ttctggacac
     3421 caagtggacc ctgctgcagg agcagggcac caagactgtg aggcagaacc tggagccgtt
     3481 gttcgagcag tacatcaaca acctcaggag gcagctggac aacatcgtgg gggaacgggg
     3541 tcgtctggac tcggagctga gaaacatgca ggacctggtg gaggacctca agaacaaata
     3601 tgaggatgaa atcaacaagc gcacagcagc agagaatgaa tttgtgactc tgaagaagga
     3661 tgtggatgct gcctacatga acaaggttga actgcaagcc aaggcagaca ctcttacaga
     3721 tgagatcaac ttcctgagag ccttgtatga tgcagagctg tcccagatgc agacccacat
     3781 ctcagacaca tccgtggtgc tatccatgga caacaaccgc aacctggacc tggacagcat
     3841 catcgctgag gtcaaggccc aatatgagga gattgctcag aggagcaggg ctgaggctga
     3901 gtcctggtac cagacaaagt acgaggagct gcagatcaca gcaggcagac atggggacga
     3961 cctgcgcaac accaagcagg agattgctga gatcaaccgc atgatccaga ggctgagatc
     4021 tgagatcgac cacgtcaaga agcagtgtgc caacctacag gccgccattg ctgatgctga
     4081 gcagcgtggg gagatggccc tcaaggatgc taagaacaag ctggaagggc tggaggatgc
     4141 cctgcagaag gccaagcagg acctggcccg gctgctgaag gagtaccagg agctgatgaa
     4201 cgtcaagctg gccctggatg tggagatcgc cacctaccgc aagctgctgg agggcgagga
     4261 gtgcaggctg aatggcgaag gcgttggaca agtcaacatc tctgtagtgc agtccaccgt
     4321 ctccagtggc tatggcggtg ccagcggtgt cggcagtggc ttaggcctgg gtggaggaag
     4381 cagctactcc tatggcagtg gtcttggcgt tggaggcggc tttagttcca gcagcggcag
     4441 agccactggg ggtggcctca gctctgttgg aggcggcagt tccaccatca agtacaccac
     4501 cacctcctcc tccagcagga agagctacaa gcacggatcc tacccatacg acgtacctga
     4561 ttatgcaact tatccttatg atgtacctga ttatgctagc tatccttatg atgtcccaga
     4621 ctacgcatag tctagataat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     4681 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     4741 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     4801 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     4861 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     4921 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     4981 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     5041 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     5101 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag
     5161 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     5221 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     5281 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     5341 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     5401 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     5461 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     5521 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     5581 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     5641 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     5701 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     5761 gcctgaccgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5821 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5881 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5941 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     6001 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     6061 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6121 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     6181 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6241 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6301 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6361 ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt
     6421 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     6481 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     6541 gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga
     6601 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag
     6661 aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta
     6721 gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta
     6781 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt
     6841 gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac
     6901 tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc
     6961 ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc
     7021 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     7081 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     7141 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     7201 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     7261 aatatcagag agtgagaggc cttgacattg ctagcgtttt accgtcgacc tctagctaga
     7321 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7381 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7441 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7501 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8161 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     8281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     8341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8401 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8461 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8521 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8581 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8641 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8701 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8761 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8821 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     8881 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8941 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     9001 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     9061 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     9121 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9181 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     9241 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9301 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9361 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9421 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     9481 tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac
     9541 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9601 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa
     9661 gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg
     9721 tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt
     9781 atttgttaaa tatgtactac aaacttagta gtttttaaag aaattgtatt tgttaaatat
     9841 gtactacaaa cttagtagt
//
