LOCUS       V022008                 8651 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022008
VERSION     V022008
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             649..1365
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1399..3363
                     /label="FOXO1(NM_002015)"
                     /note="FOXO1(NM_002015)"
                     /gene="FOXO1"
     misc_feature    3427..4015
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4081..4129
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4331..4759
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4773..5102
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5169..5963
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6139..6260
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6309..6325)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6333..6349
                     /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(6357..6387)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6402..6423
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6711..7299)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7470..8330)
                     /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(8331..8435)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgccaccat ggtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc
     1381 ggtaccgagc tcggatccat ggccgaggcg cctcaggtgg tggagatcga cccggacttc
     1441 gagccgctgc cccggccgcg ctcgtgcacc tggccgctgc ccaggccgga gtttagccag
     1501 tccaactcgg ccacctccag cccggcgccg tcgggcagcg cggctgccaa ccccgacgcc
     1561 gcggcgggcc tgccctcggc ctcggctgcc gctgtcagcg ccgacttcat gagcaacctg
     1621 agcttgctgg aggagagcga ggacttcccg caggcgcccg gctccgtggc ggcggcggtg
     1681 gcggcggcgg ccgccgcggc cgccaccggg gggctgtgcg gggacttcca gggcccggag
     1741 gcgggctgcc tgcacccagc gccaccgcag cccccgccgc ccgggccgct gtcgcagcac
     1801 ccgccggtgc cccccgccgc cgctgggccg ctcgcggggc agccgcgcaa gagcagctcg
     1861 tcccgccgca acgcgtgggg caacctgtcc tacgccgacc tcatcaccaa ggccatcgag
     1921 agctcggcgg agaagcggct cacgctgtcg cagatctacg agtggatggt caagagcgtg
     1981 ccctacttca aggataaggg tgacagcaac agctcggcgg gctggaagaa ttcaattcgt
     2041 cataatctgt ccctacacag caagttcatt cgtgtgcaga atgaaggaac tggaaaaagt
     2101 tcttggtgga tgctcaatcc agagggtggc aagagcggga aatctcctag gagaagagct
     2161 gcatccatgg acaacaacag taaatttgct aagagccgaa gccgagctgc caagaagaaa
     2221 gcatctctcc agtctggcca ggagggtgct ggggacagcc ctggatcaca gttttccaaa
     2281 tggcctgcaa gccctggctc tcacagcaat gatgactttg ataactggag tacatttcgc
     2341 cctcgaacta gctcaaatgc tagtactatt agtgggagac tctcacccat tatgaccgaa
     2401 caggatgatc ttggagaagg ggatgtgcat tctatggtgt acccgccatc tgccgcaaag
     2461 atggcctcta ctttacccag tctgtctgag ataagcaatc ccgaaaacat ggaaaatctt
     2521 ttggataatc tcaaccttct ctcatcacca acatcattaa ctgtttcgac ccagtcctca
     2581 cctggcacca tgatgcagca gacgccgtgc tactcgtttg cgccaccaaa caccagtttg
     2641 aattcaccca gcccaaacta ccaaaaatat acatatggcc aatccagcat gagccctttg
     2701 ccccagatgc ctatacaaac acttcaggac aataagtcga gttatggagg tatgagtcag
     2761 tataactgtg cgcctggact cttgaaggag ttgctgactt ctgactctcc tccccataat
     2821 gacattatga caccagttga tcctggggta gcccagccca acagccgggt tctgggccag
     2881 aacgtcatga tgggccctaa ttcggtcatg tcaacctatg gcagccaggc atctcataac
     2941 aaaatgatga atcccagctc ccatacccac cctggacatg ctcagcagac atctgcagtt
     3001 aacgggcgtc ccctgcccca cacggtaagc accatgcccc acacctcggg tatgaaccgc
     3061 ctgacccaag tgaagacacc tgtacaagtg cctctgcccc accccatgca gatgagtgcc
     3121 ctggggggct actcctccgt gagcagctgc aatggctatg gcagaatggg ccttctccac
     3181 caggagaagc tcccaagtga cttggatggc atgttcattg agcgcttaga ctgtgacatg
     3241 gaatccatca ttcggaatga cctcatggat ggagatacat tggattttaa ctttgacaat
     3301 gtgttgccca accaaagctt cccacacagt gtcaagacaa cgacacatag ctgggtgtca
     3361 ggctgaattc tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgatat
     3421 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3481 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3541 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3601 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3661 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3721 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3781 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     3841 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3901 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3961 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     4021 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4081 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     4141 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4201 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4261 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     4321 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4381 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4441 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     4501 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4561 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4621 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4681 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     4741 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4801 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4861 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4921 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4981 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5041 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5101 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     5161 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5221 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5281 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5341 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5401 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5461 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5521 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5581 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5641 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     5701 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5761 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5821 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5881 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5941 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     6001 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6061 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6121 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6181 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6241 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6301 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6361 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6421 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6481 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6541 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6601 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6661 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6721 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6781 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6841 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6901 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6961 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7021 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7081 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7141 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7201 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7261 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7321 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7381 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7441 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7501 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7561 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7621 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7681 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7741 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7801 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7861 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7921 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7981 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8041 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8101 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8161 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8221 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8281 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8341 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8401 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8461 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     8521 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     8581 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     8641 atctgcttag g
//
