LOCUS       V042536                 9901 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042536
VERSION     V042536
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2284..2991
                     /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"
     CDS             3028..4992
                     /label="FOXO1(NM_002015)"
                     /note="FOXO1(NM_002015)"
                     /gene="FOXO1"
     CDS             4999..5025
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        5066..5277
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5290..5558
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5591..5989
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5996..6584
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6658..6891
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6963..7084
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7103..7238
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7276..7292)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7300..7316
                     /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(7324..7354)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7369..7390
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7678..8266)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8437..9297)
                     /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(9298..9402)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9876..9892
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagatgcc
     2281 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     2341 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2401 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2461 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2521 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2581 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2641 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2701 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2761 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2821 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2881 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2941 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gggaggtgga
     3001 ggtagtggag gaggcggatc gtctagaatg gccgaggcgc ctcaggtggt ggagatcgac
     3061 ccggacttcg agccgctgcc ccggccgcgc tcgtgcacct ggccgctgcc caggccggag
     3121 tttagccagt ccaactcggc cacctccagc ccggcgccgt cgggcagcgc ggctgccaac
     3181 cccgacgccg cggcgggcct gccctcggcc tcggctgccg ctgtcagcgc cgacttcatg
     3241 agcaacctga gcttgctgga ggagagcgag gacttcccgc aggcgcccgg ctccgtggcg
     3301 gcggcggtgg cggcggcggc cgccgcggcc gccaccgggg ggctgtgcgg ggacttccag
     3361 ggcccggagg cgggctgcct gcacccagcg ccaccgcagc ccccgccgcc cgggccgctg
     3421 tcgcagcacc cgccggtgcc ccccgccgcc gctgggccgc tcgcggggca gccgcgcaag
     3481 agcagctcgt cccgccgcaa cgcgtggggc aacctgtcct acgccgacct catcaccaag
     3541 gccatcgaga gctcggcgga gaagcggctc acgctgtcgc agatctacga gtggatggtc
     3601 aagagcgtgc cctacttcaa ggataagggt gacagcaaca gctcggcggg ctggaagaat
     3661 tcaattcgtc ataatctgtc cctacacagc aagttcattc gtgtgcagaa tgaaggaact
     3721 ggaaaaagtt cttggtggat gctcaatcca gagggtggca agagcgggaa atctcctagg
     3781 agaagagctg catccatgga caacaacagt aaatttgcta agagccgaag ccgagctgcc
     3841 aagaagaaag catctctcca gtctggccag gagggtgctg gggacagccc tggatcacag
     3901 ttttccaaat ggcctgcaag ccctggctct cacagcaatg atgactttga taactggagt
     3961 acatttcgcc ctcgaactag ctcaaatgct agtactatta gtgggagact ctcacccatt
     4021 atgaccgaac aggatgatct tggagaaggg gatgtgcatt ctatggtgta cccgccatct
     4081 gccgcaaaga tggcctctac tttacccagt ctgtctgaga taagcaatcc cgaaaacatg
     4141 gaaaatcttt tggataatct caaccttctc tcatcaccaa catcattaac tgtttcgacc
     4201 cagtcctcac ctggcaccat gatgcagcag acgccgtgct actcgtttgc gccaccaaac
     4261 accagtttga attcacccag cccaaactac caaaaatata catatggcca atccagcatg
     4321 agccctttgc cccagatgcc tatacaaaca cttcaggaca ataagtcgag ttatggaggt
     4381 atgagtcagt ataactgtgc gcctggactc ttgaaggagt tgctgacttc tgactctcct
     4441 ccccataatg acattatgac accagttgat cctggggtag cccagcccaa cagccgggtt
     4501 ctgggccaga acgtcatgat gggccctaat tcggtcatgt caacctatgg cagccaggca
     4561 tctcataaca aaatgatgaa tcccagctcc catacccacc ctggacatgc tcagcagaca
     4621 tctgcagtta acgggcgtcc cctgccccac acggtaagca ccatgcccca cacctcgggt
     4681 atgaaccgcc tgacccaagt gaagacacct gtacaagtgc ctctgcccca ccccatgcag
     4741 atgagtgccc tggggggcta ctcctccgtg agcagctgca atggctatgg cagaatgggc
     4801 cttctccacc aggagaagct cccaagtgac ttggatggca tgttcattga gcgcttagac
     4861 tgtgacatgg aatccatcat tcggaatgac ctcatggatg gagatacatt ggattttaac
     4921 tttgacaatg tgttgcccaa ccaaagcttc ccacacagtg tcaagacaac gacacatagc
     4981 tgggtgtcag gcctcgagta cccatacgac gtaccagatt acgcttgaat tcgaaggatc
     5041 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     5101 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     5161 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     5221 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     5281 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     5341 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     5401 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     5461 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     5521 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg
     5581 ctaggccacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     5641 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     5701 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     5761 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     5821 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     5881 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     5941 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag tcgacaatca
     6001 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6061 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6121 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6181 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6241 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6301 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6361 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     6421 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6481 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6541 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta cctttaagac
     6601 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6661 aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct
     6721 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6781 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6841 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     6901 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     6961 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7021 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7081 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc
     7141 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     7201 gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc
     7261 aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7321 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7381 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7441 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7501 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7561 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7621 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7681 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7741 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7801 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7861 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7921 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7981 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8041 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8101 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8161 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8221 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8281 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8341 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8401 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8521 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8581 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8641 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8701 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8761 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8821 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8881 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8941 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9001 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9061 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9121 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9181 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9241 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9301 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9361 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9421 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     9481 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     9541 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     9601 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     9661 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     9721 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     9781 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     9841 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     9901 g
//
