LOCUS       V024534                 9352 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024534
VERSION     V024534
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..4048
                     /label="Ager(NM_007425)"
                     /note="Ager(NM_007425)"
                     /gene="Ager"
     CDS             4055..4120
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4140..4639
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4660..5259
                     /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    5273..5861
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6068..6701
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6830..6846)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6854..6870
                     /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(6878..6908)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6923..6944
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7232..7820)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7991..8851)
                     /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(8852..8956)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9004..9138
                     /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 ccatgccagc ggggacagca gctagagcct gggtgctggt
     2881 tcttgctcta tggggagctg tagctggtgg tcagaacatc acagcccgga ttggagagcc
     2941 acttgtgcta agctgtaagg gggcccctaa gaagccgccc cagcagctag aatggaaact
     3001 gaacacagga agaactgaag cttggaaggt cctctctccc cagggaggcc cctgggacag
     3061 cgtggctcga atcctcccca atggttccct cctccttcca gccactggaa ttgtcgatga
     3121 ggggactttc cggtgtcggg caactaacag gcgagggaag gaggtcaagt ccaactaccg
     3181 agtccgagtc taccagattc ctgggaagcc agaaattgtg gatcctgcct ctgaactcac
     3241 agccagtgtc cctaataagg tggggacatg tgtgtctgag ggaagctacc ctgcagggac
     3301 ccttagctgg cacttagatg ggaaacttct gattcccgat ggcaaagaaa cactcgtgaa
     3361 ggaagagacc aggagacacc ctgagacggg actctttaca ctgcggtcag agctgacagt
     3421 gatccccacc caaggaggaa cccatcctac cttctcctgc agtttcagcc tgggccttcc
     3481 ccggcgcaga cccctgaaca cagcccccat ccaactccga gtcagggagc ctgggcctcc
     3541 agagggcatt cagctgttgg ttgagcctga aggtggaata gtcgctcctg gtgggactgt
     3601 gaccttgacc tgtgccatct ctgcccagcc ccctcctcag gtccactgga taaaggatgg
     3661 tgcacccttg cccctggctc ccagccctgt gctgctcctc cctgaggtgg ggcacgagga
     3721 tgagggcacc tatagctgcg tggccaccca ccctagccac ggacctcagg aaagccctcc
     3781 tgtcagcatc agggtcacag aaaccggcga tgaggggcca gctgaaggct ctgtgggtga
     3841 gtctgggctg ggtacgctag ccctggcctt ggggatcctg ggaggcctgg gagtagtagc
     3901 cctgctcgtc ggggctatcc tgtggcgaaa acgacaaccc aggcgtgagg agaggaaggc
     3961 cccggaaagc caggaggatg aggaggaacg tgcagagctg aatcagtcag aggaagcgga
     4021 gatgccagag aatggtgccg ggggaccggg atccgactac aaagaccatg acggtgatta
     4081 taaagatcat gacatcgact acaaggatga cgatgacaag tagtctagat aattctaccg
     4141 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     4201 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     4261 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     4321 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     4381 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     4441 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     4501 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     4561 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     4621 ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg
     4681 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     4741 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     4801 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4861 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4921 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4981 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     5041 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     5101 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     5161 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     5221 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc
     5281 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5341 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5401 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5461 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5521 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5581 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5641 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     5701 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5761 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     5821 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt
     5881 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
     5941 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta
     6001 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     6061 aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc
     6121 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga
     6181 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa
     6241 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat
     6301 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg
     6361 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga
     6421 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc
     6481 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca
     6541 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     6601 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     6661 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     6721 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca
     6781 ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6841 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6961 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7021 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7801 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7861 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7921 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7981 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8041 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8101 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8161 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8221 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8281 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8341 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8401 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8461 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8521 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8581 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8641 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8701 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8761 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8821 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8881 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8941 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     9001 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     9061 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     9121 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
     9181 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
     9241 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
     9301 gtagttttta aagaaattgt atttgttaaa tatgtactac aaacttagta gt
//
