LOCUS       V038026                 9676 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038026
VERSION     V038026
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..4372
                     /label="PIAS4(NM_015897)"
                     /note="PIAS4(NM_015897)"
                     /gene="PIAS4"
     CDS             4379..4444
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4464..4963
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4984..5583
                     /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    5597..6185
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6392..7025
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7154..7170)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7178..7194
                     /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(7202..7232)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7247..7268
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7556..8144)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8315..9175)
                     /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(9176..9280)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9328..9462
                     /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 ccatggcggc ggagctggtg gaggccaaaa acatggtgat
     2881 gagttttcga gtctccgacc ttcagatgct cctgggtttc gtgggccgga gtaagagtgg
     2941 actgaagcac gagctcgtca ccagggccct ccagctggtg cagtttgact gtagccctga
     3001 gctgttcaag aagatcaagg agctgtacga gacccgctac gccaagaaga actcggagcc
     3061 tgccccacag ccgcaccggc ccctggaccc cctgaccatg cactccacct acgaccgggc
     3121 cggcgctgtg cccaggactc cgctggcagg ccccaatatt gactaccccg tgctctacgg
     3181 aaagtactta aacggactgg gacggttgcc cgccaagacc ctcaagccag aagtccgcct
     3241 ggtgaagctg ccgttcttta atatgctgga tgagctgctg aagcccaccg aattagtccc
     3301 acagaacaac gagaagcttc aggagagccc gtgcatcttc gcattgacgc caagacaggt
     3361 ggagttgatc cggaactcca gggaactgca gcccggagtt aaagccgtgc aggtcgtcct
     3421 gagaatctgt tactcagaca ccagctgccc tcaggaggac cagtacccgc ccaacatcgc
     3481 tgtgaaggtc aaccacagct actgctccgt cccgggctac tacccctcca ataagcccgg
     3541 ggtggagccc aagaggccgt gccgccccat caacctcacc cacctcatgt acctgtcctc
     3601 ggccaccaac cgcatcactg tcacctgggg gaactacggc aagagctact cggtggccct
     3661 gtacctggtg cggcagctga cctcatcgga gctgctgcag aggctgaaga ccattggggt
     3721 aaagcacccg gagctgtgca aggcactggt caaggagaag ctgcgccttg atcctgacag
     3781 cgagatcgcc accaccggtg tgcgggtgtc cctcatctgt ccgctggtga agatgcggct
     3841 ctccgtgccc tgccgggcag agacctgtgc ccacctgcag tgcttcgacg ccgtcttcta
     3901 cctgcagatg aacgagaaga agcccacctg gatgtgcccc gtgtgcgaca agccagcccc
     3961 ctacgaccag ctcatcatcg acgggctcct ctcgaagatc ctgagcgagt gtgaggacgc
     4021 cgacgagatc gagtacctgg tggacggctc gtggtgcccg atccgcgccg aaaaggagcg
     4081 cagctgcagc ccgcagggcg ccatcctcgt gctgggcccc tcggacgcca atgggctcct
     4141 gcccgccccc agcgtcaacg ggagcggtgc cctgggcagc acgggtggcg gcggcccggt
     4201 gggcagcatg gagaatggga agccgggcgc cgatgtggtg gacctcacgc tggacagctc
     4261 atcgtcctcg gaggatgagg aggaggagga agaggaggag gaagacgagg acgaagaggg
     4321 gccccggccc aagcgccgct gccccttcca gaagggcctg gtgccggcct gcggatccga
     4381 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     4441 caagtagtct agataattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc
     4501 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac
     4561 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc
     4621 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
     4681 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
     4741 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
     4801 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
     4861 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
     4921 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt
     4981 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     5041 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     5101 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     5161 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     5221 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     5281 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     5341 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     5401 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     5461 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     5521 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     5581 tgaccgcgtc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5641 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5701 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5761 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5821 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     5881 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     5941 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     6001 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6061 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6121 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6181 cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc
     6241 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     6301 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     6361 cttagccact ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga
     6421 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
     6481 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
     6541 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
     6601 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
     6661 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
     6721 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
     6781 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
     6841 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     6901 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     6961 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7021 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     7081 atcagagagt gagaggcctt gacattgcta gcgttttacc gtcgacctct agctagagct
     7141 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7201 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7261 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7321 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7381 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7441 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7501 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7561 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7621 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7681 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7741 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7801 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7861 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7921 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7981 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     8041 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8101 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8161 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8221 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8281 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8341 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8401 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8461 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8521 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8581 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8641 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8701 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8761 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8821 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8881 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8941 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9001 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9061 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9121 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9181 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9241 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9301 cacctgacgt cgacggatcg ggagatcaac ttgtttattg cagcttataa tggttacaaa
     9361 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     9421 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct
     9481 aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg
     9541 tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt
     9601 tgttaaatat gtactacaaa cttagtagtt tttaaagaaa ttgtatttgt taaatatgta
     9661 ctacaaactt agtagt
//
