LOCUS       V034703                 9856 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034703
VERSION     V034703
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..4552
                     /label="Nfib(NM_001113209)"
                     /note="Nfib(NM_001113209)"
                     /gene="Nfib"
     CDS             4559..4624
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4644..5143
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5164..5763
                     /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    5777..6365
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6572..7205
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7334..7350)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7358..7374
                     /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(7382..7412)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7427..7448
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7736..8324)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8495..9355)
                     /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(9356..9460)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9508..9642
                     /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 ccatgatgta ttctcccatc tgtctcactc aggatgaatt
     2881 ccacccattt attgaggcac ttcttcctca cgtccgtgca attgcctata cttggttcaa
     2941 cctgcaggct cgaaaacgca agtactttaa aaagcatgag aaacgaatgt cgaaggatga
     3001 agaaagggca gtcaaagacg agctgctcag tgagaagccc gaaatcaagc agaagtgggc
     3061 atccaggctc ctggccaaac tgcgcaaaga tatccgccag gagtaccggg aggactttgt
     3121 gcttaccgtg actggcaaga agcacccgtg ctgtgtctta tccaatccag accagaaggg
     3181 taagattagg aggatcgact gcctgcgaca ggcagacaaa gtctggcgtc tggatctagt
     3241 catggtgatc ctgttcaaag gcatcccttt ggagagtacg gatggagagc gactcatgaa
     3301 gtccccgcac tgcacaaacc cagcactttg tgttcagcca caccacatca cagtatcagt
     3361 taaggagctt gacttgtttt tggcatacta cgtgcaggag caagattctg gacaatcagg
     3421 aagtccaagc cacagtgatc ctgccaagaa tcctccaggg tacctcgagg acagctttgt
     3481 aaaatccgga gtcttcaatg tatcagagct tgtgagagta tccagaacac ccataaccca
     3541 gggaactgga gtcaacttcc caatcggaga aattcccagc caaccatact atcatgacat
     3601 gaactctggt gtgaacctgc agaggtcgct gtcttctcca ccgagcagca aaagacccaa
     3661 aactatatct atagatgaaa atatggagcc aagtcctaca ggagactttt acccctctcc
     3721 aaattcacca gctgctggaa gtcgaacatg gcatgaacga gatcaagata tgtcttctcc
     3781 aactacaatg aagaagcctg agaagccact gtttagctct acatctccac aggattcttc
     3841 cccaagattg agcactttcc cccagcacca tcatcccgga atacctggag tcgcgcacag
     3901 tgtcatctca actcgaactc cacctccgcc ctcaccgttg ccatttccga cgcaagctat
     3961 ccttcctccg gcaccttcca gctacttctc tcatccaaca atcagatatc ctcctcacct
     4021 gaatcctcag gatactctga agaactacgt accttcttat gacccatcca gtcctcaaac
     4081 gagccagcct aacagcagtg gtcaagtagt agggaaagtg cctggccatt tcacacctgt
     4141 cttggcaccc tctccccatc ccagtgcagt gcgacctgtg accctgacca tgacagatac
     4201 taaacccatc actacatcca ctgaaggtga ggcagcttca cctacagcaa ccacctacac
     4261 agcctcaggc acatctcaag ccaatcgata tgtgggacta agcccaagag acccatcctt
     4321 cctgcatcag caacagctga ggatttgtga ctggaccatg aatcaaaacg gcaggcattt
     4381 ataccccagt accagtgagg atacattggg aattacttgg caaagtcctg gtacctgggc
     4441 tagcttggtt ccttttcaag tgtcaaatag gacacccatc ttaccggcaa atgtccaaaa
     4501 ttatggtttg aacataattg gagagccttt ccttcaagcg gagacaagca acggatccga
     4561 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     4621 caagtagtct agataattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc
     4681 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac
     4741 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc
     4801 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
     4861 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
     4921 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
     4981 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
     5041 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
     5101 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt
     5161 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     5221 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     5281 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     5341 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     5401 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     5461 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     5521 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     5581 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     5641 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     5701 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     5761 tgaccgcgtc tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5821 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5881 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5941 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6001 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6061 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6121 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     6181 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6241 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6301 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6361 cccgcctgga attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc
     6421 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     6481 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     6541 cttagccact ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga
     6601 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
     6661 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
     6721 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
     6781 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
     6841 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
     6901 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
     6961 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
     7021 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7081 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7141 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7201 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     7261 atcagagagt gagaggcctt gacattgcta gcgttttacc gtcgacctct agctagagct
     7321 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7381 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7441 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7501 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7561 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7621 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7681 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7741 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7801 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7861 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7921 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7981 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     8041 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     8101 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     8161 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     8221 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8281 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8341 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8401 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8461 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8521 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8581 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8641 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8701 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8761 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8821 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8881 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8941 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     9001 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9061 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9121 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9181 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9241 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9301 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9361 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9421 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9481 cacctgacgt cgacggatcg ggagatcaac ttgtttattg cagcttataa tggttacaaa
     9541 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     9601 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct
     9661 aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg
     9721 tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt
     9781 tgttaaatat gtactacaaa cttagtagtt tttaaagaaa ttgtatttgt taaatatgta
     9841 ctacaaactt agtagt
//
