LOCUS       V027531                 7461 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027531
VERSION     V027531
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             927..2264
                     /label="Carm1(NM_001411233)"
                     /note="Carm1(NM_001411233)"
                     /gene="Carm1"
     CDS             2304..3023
                     /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"
     polyA_signal    3061..3285
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3331..3759
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3773..4102
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4169..4963
                     /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    5137..5258
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5307..5323)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5331..5347
                     /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(5355..5385)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5400..5421
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5709..6294)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6465..7325)
                     /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(7326..7430)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaagcc actgggatcc gccaccatga tgcaggacta tgtgcggaca ggcacctacc
      961 agcgtgcgat cctgcagaac cacacggact tcaaggacaa gatcgttcta gatgtgggct
     1021 gtggctctgg gatcctgtca ttttttgctg ctcaagcagg agccaggaaa atttatgcag
     1081 tggaagccag caccatggct cagcatgcag aggtcctggt gaagagtaac aatctgacag
     1141 accgcatcgt ggtcatccct ggcaaagtag aggaggtctc attgcctgag caagtggaca
     1201 ttatcatctc agagcccatg ggctacatgc tcttcaatga acgaatgctc gagagctacc
     1261 tccatgccaa aaagtacctg aagcctagtg gaaacatgtt ccccaccatt ggtgatgtcc
     1321 acctcgcacc cttcactgat gaacagctct acatggagca gttcaccaaa gccaacttct
     1381 ggtaccagcc atccttccat ggagtggacc tgtcggccct cagaggcgcc gctgtggatg
     1441 agtacttccg gcaacctgtg gtggacacat ttgacatccg gatcctgatg gccaaatctg
     1501 tcaagtacac agtgaacttc ttagaagcca aagaaggcga tttgcacagg atagaaatcc
     1561 cattcaaatt ccacatgctg cattcagggc tagtccatgg cttggccttc tggttcgatg
     1621 ttgctttcat tggctccata atgaccgtgt ggctatccac agccccaaca gagcccctga
     1681 cccactggta ccaggtccgg tgcctcttcc agtcaccgtt gtttgccaag gccggggaca
     1741 cgctctcagg gacatgtctg cttattgcca acaaaagaca gagctatgac atcagtattg
     1801 tggcacaggt ggaccagaca ggctccaagt ccagtaacct gctggatcta aagaacccct
     1861 tcttcaggta cacaggtaca accccatcac ccccacctgg ctcacactac acgtctccct
     1921 cggagaatat gtggaacaca ggaagcacct ataatctcag cagcggggtg gctgtggctg
     1981 gaatgcctac tgcctacgac ctgagcagtg ttattgccgg cggctccagt gtgggtcaca
     2041 acaacctgat tcccttagct aacacaggga ttgtcaatca cacccactcc cggatgggct
     2101 ccataatgag cacgggcatt gtccaaggct cctcaggtgc ccagggaggc ggcggtagct
     2161 ccagtgccca ctatgcagtc aacaaccagt tcaccatggg tggccctgcc atctctatgg
     2221 cctcgcccat gtccatcccg accaacacca tgcactatgg gagtggctcg agtggcggtg
     2281 gaggaagcgg cggaggtgga tctatggtga gcaagggcga ggagctgttc accggggtgg
     2341 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2401 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2461 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2521 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2581 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2641 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2701 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2761 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2821 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2881 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2941 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     3001 gcatggacga gctgtacaag tagtctagag ggcccgttta aacccgctga tcagcctcga
     3061 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3121 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3181 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3241 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3301 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3361 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3421 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3481 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3541 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3601 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3661 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3721 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3781 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3841 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3901 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3961 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4021 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4081 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4141 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4201 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4261 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4321 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4381 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4441 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4501 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4561 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4621 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4681 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4741 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4801 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4861 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4921 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4981 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5041 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5101 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5161 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5221 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5281 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5341 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5401 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5461 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5521 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5581 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5641 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5701 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5761 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5821 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5881 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5941 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6001 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6061 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6121 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6181 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6241 agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6301 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6361 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6421 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6481 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6541 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6601 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6661 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6721 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6781 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6841 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6901 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6961 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7021 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7081 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7141 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7201 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7261 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7321 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7381 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7441 cgaaaagtgc cacctgacgt c
//
