LOCUS       V046156                 9780 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046156
VERSION     V046156
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..3829
                     /label="Foxs1(NM_010226)"
                     /note="Foxs1(NM_010226)"
                     /gene="Foxs1"
     CDS             3878..4588
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     promoter        4615..5114
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5135..5734
                     /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    5748..6336
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6543..7176
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7305..7321)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7329..7345
                     /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(7353..7383)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7398..7419
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7707..8295)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8466..9326)
                     /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(9327..9431)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9479..9613
                     /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 ccatgcagaa gcaaccatca ccagaatcct tggctccttc
     2881 tgcagagccc accaagcccc cttacagcta catagccctg attgccatgg ctatccagag
     2941 ttcaccgggt cagcgggcca ccctcagtgg catctaccgc tacatcatgg gccgctttgc
     3001 cttttaccgg cacaaccggc cgggctggca aaacagcatc cgccataacc tgtctctcaa
     3061 cgagtgtttc gtcaaggtgc ctcgtgatga ccgcaagcca ggcaagggca gctactggac
     3121 cctggatccg gactgccacg acatgttcca gcatggcagc tttctccgtc gccgcagacg
     3181 cttcaccaaa cgcacaggtg cgcagggcac caagggccct gtcaagatag accacagacc
     3241 ccacagagcc accagcccgg acccaggagc ccccaagacc acaactggca gactgtgccc
     3301 attcccacaa gaggtgccaa accccaaggg cttaagcttt gagggtctga tggggtcttt
     3361 gccagccaac atgagctcaa caaccagtga cgtcaggccc caactgccca ctggacccaa
     3421 agagatgtgc tcggcaaagt ctggaggccc aagggagctc tctgaagcta cctcgccctc
     3481 accgtgccca gcattcggct tttcatctgc cttctctgac gctgagagtc tcggcaaggc
     3541 ccccacaccc ggtgtggctc ctgagagcgt tgggagcagc taccagtgcc ggatgcagac
     3601 actgaatttc tgtatgggga ctgacccagg ccttgaacac ctcctggtct cttcagtccc
     3661 cacccctgga tcctcgacac cttcagcctc tcaccgggcc ccactgcccc tgccggctga
     3721 ctccaaagaa ccctgggtgg ctgggagctt ccctgtccag ggaggttctg gctatccact
     3781 gggattaccc ccctgtcttt accggactcc gggaatgttc ttcttcgagg gtggaggcgg
     3841 atcaggaggc ggtggatctg atccaccggt cgccaccatg gtgagcaagg gcgaggagga
     3901 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     3961 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     4021 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     4081 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     4141 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     4201 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     4261 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     4321 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     4381 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     4441 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     4501 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     4561 cggcggcatg gacgagctgt acaagtagcg gccgcgactc tagataattc taccgggtag
     4621 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
     4681 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
     4741 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     4801 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     4861 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     4921 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     4981 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     5041 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     5101 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg
     5161 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     5221 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     5281 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     5341 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5401 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5461 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5521 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5581 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5641 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5701 cacctggtgc atgacccgca agcccggtgc ctgaccgcgt ctggaacaat caacctctgg
     5761 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5821 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5881 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5941 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     6001 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6061 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6121 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     6181 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6241 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6301 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga
     6361 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     6421 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     6481 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg
     6541 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     6601 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     6661 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     6721 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     6781 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     6841 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     6901 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     6961 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     7021 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     7081 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     7141 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     7201 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct
     7261 agcgttttac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     7321 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7381 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7441 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7501 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7561 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7621 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8401 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8461 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8521 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8641 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8761 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8881 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     9001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9061 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     9121 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9301 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9361 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9421 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa
     9481 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     9541 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     9601 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     9661 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
     9721 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
