LOCUS       V027844                 7909 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027844
VERSION     V027844
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /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             1642..3399
                     /label="ACSM3(NM_005622)"
                     /note="ACSM3(NM_005622)"
                     /gene="ACSM3"
     polyA_signal    3476..3700
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3746..4174
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4188..4517
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4584..5378
                     /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    5552..5673
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5722..5738)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5746..5762
                     /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(5770..5800)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5815..5836
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6124..6709)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6880..7740)
                     /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(7741..7845)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catgctagct cgtgtcacca ggaagatgct acgtcatgcc
     1681 aagtgttttc agcgcctagc aatttttggt tctgtgaggg cactgcataa agataataga
     1741 acagcaaccc ctcagaattt ctccaactat gaatccatga aacaggactt caaactgggg
     1801 attccagagt atttcaactt tgctaaagat gtcctggacc aatggactga taaggaaaag
     1861 gctggaaaga aaccttcaaa tccagccttc tggtggatca acagaaatgg agaagagatg
     1921 cgatggagtt ttgaggaact gggatctctg tccagaaaat ttgccaatat actttcagaa
     1981 gcctgttccc tacaaagagg agatcgggta attctgattc tgcccagggt cccagagtgg
     2041 tggcttgcaa atgtggcctg tctgcgaaca gggacagttt taattccagg aaccactcag
     2101 ctgacccaga aagacattct ctacagacta caatcttcaa aagcaaactg cattatcacc
     2161 aatgatgttt tagccccagc agtagacgct gttgcatcca aatgtgaaaa tctgcactcc
     2221 aagctgattg tatcagagaa ctccagagag gggtggggga acctcaagga gttgatgaaa
     2281 catgccagtg acagccacac ctgtgtgaag acaaaacaca atgagatcat ggccatattc
     2341 tttaccagtg gaacaagtgg atatccgaaa atgactgcac acacccacag cagttttggt
     2401 ttaggattat ctgtaaatgg aaggttctgg ctagatttga caccctcaga tgtgatgtgg
     2461 aatacctcag atacgggctg ggcaaagtct gcatggagta gtgttttttc tccgtggatc
     2521 cagggagcat gtgtattcac acaccattta ccccgttttg agccgacttc tatcttgcaa
     2581 acactctcca agtaccccat cacagtcttc tgttcagcac caactgtata ccgaatgctt
     2641 gtacagaatg atataaccag ctataagttt aaaagcttaa agcactgtgt gagtgctggg
     2701 gaaccaatta cccctgacgt gactgaaaaa tggagaaaca agacgggcct ggatatctac
     2761 gaaggatatg gacagactga aacggtgcta atctgtggaa attttaaggg aatgaaaatt
     2821 aaacctggct caatgggaaa accttctcct gctttcgatg ttaagattgt agatgtaaat
     2881 ggcaatgttc tacctcctgg acaagaagga gatattggca ttcaagttct acccaaccga
     2941 ccatttggcc tttttactca ttacgtagat aatccttcaa aaacagcttc aactctacga
     3001 ggcaatttct atatcactgg ggacagagga tatatggata aagatgggta tttctggttt
     3061 gttgcaagag cagatgatgt catattatcc tctggctatc gaattggacc atttgaggta
     3121 gaaaatgccc tgaatgaaca cccttcagtt gcagagtcag ctgttgtcag cagcccagac
     3181 cccatcagag gagaggtagt aaaggctttt gtcgttctaa atcctgatta caagtcacat
     3241 gatcaagaac aactaataaa ggagattcag gagcatgtta aaaaaactac agcaccttac
     3301 aaatatccca gaaaggtaga atttattcaa gagctgccaa agactatcag tgggaagaca
     3361 aaaagaaatg aactgaggaa gaaagaatgg aagacaattt aaggatcctg atatccagca
     3421 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     3481 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3541 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3601 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3661 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3721 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3781 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3841 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3901 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3961 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4021 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4081 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4141 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4261 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4381 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4441 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4501 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4561 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4621 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4681 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4741 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4801 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4861 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4921 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4981 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5041 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5101 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5161 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5221 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5281 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5341 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5401 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5461 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5521 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5581 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5641 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5701 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5761 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5821 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5881 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5941 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6001 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6061 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6121 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6181 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6241 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6301 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6361 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6421 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6481 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6541 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6601 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6661 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6721 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6781 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6841 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6901 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6961 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7021 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7081 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7141 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7201 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7261 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7321 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7381 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7441 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7501 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7561 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7621 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7681 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7741 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7801 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7861 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
