LOCUS       V023902                 7599 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023902
VERSION     V023902
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             1660..3063
                     /label="IL6R(NM_000565)"
                     /note="IL6R(NM_000565)"
                     /gene="IL6R"
     polyA_signal    3166..3390
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3436..3864
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3878..4207
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4274..5068
                     /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    5242..5363
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5412..5428)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5436..5452
                     /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(5460..5490)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5505..5526
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5814..6399)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6570..7430)
                     /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(7431..7535)
                     /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 tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagctta tgctggccgt cggctgcgcg
     1681 ctgctggctg ccctgctggc cgcgccggga gcggcgctgg ccccaaggcg ctgccctgcg
     1741 caggaggtgg cgagaggcgt gctgaccagt ctgccaggag acagcgtgac tctgacctgc
     1801 ccgggggtag agccggaaga caatgccact gttcactggg tgctcaggaa gccggctgca
     1861 ggctcccacc ccagcagatg ggctggcatg ggaaggaggc tgctgctgag gtcggtgcag
     1921 ctccacgact ctggaaacta ttcatgctac cgggccggcc gcccagctgg gactgtgcac
     1981 ttgctggtgg atgttccccc cgaggagccc cagctctcct gcttccggaa gagccccctc
     2041 agcaatgttg tttgtgagtg gggtcctcgg agcaccccat ccctgacgac aaaggctgtg
     2101 ctcttggtga ggaagtttca gaacagtccg gccgaagact tccaggagcc gtgccagtat
     2161 tcccaggagt cccagaagtt ctcctgccag ttagcagtcc cggagggaga cagctctttc
     2221 tacatagtgt ccatgtgcgt cgccagtagt gtcgggagca agttcagcaa aactcaaacc
     2281 tttcagggtt gtggaatctt gcagcctgat ccgcctgcca acatcacagt cactgccgtg
     2341 gccagaaacc cccgctggct cagtgtcacc tggcaagacc cccactcctg gaactcatct
     2401 ttctacagac tacggtttga gctcagatat cgggctgaac ggtcaaagac attcacaaca
     2461 tggatggtca aggacctcca gcatcactgt gtcatccacg acgcctggag cggcctgagg
     2521 cacgtggtgc agcttcgtgc ccaggaggag ttcgggcaag gcgagtggag cgagtggagc
     2581 ccggaggcca tgggcacgcc ttggacagaa tccaggagtc ctccagctga gaacgaggtg
     2641 tccaccccca tgcaggcact tactactaat aaagacgatg ataatattct cttcagagat
     2701 tctgcaaatg cgacaagcct cccagtgcaa gattcttctt cagtaccact gcccacattc
     2761 ctggttgctg gagggagcct ggccttcgga acgctcctct gcattgccat tgttctgagg
     2821 ttcaagaaga cgtggaagct gcgggctctg aaggaaggca agacaagcat gcatccgccg
     2881 tactctttgg ggcagctggt cccggagagg cctcgaccca ccccagtgct tgttcctctc
     2941 atctccccac cggtgtcccc cagcagcctg gggtctgaca atacctcgag ccacaaccga
     3001 ccagatgcca gggacccacg gagcccttat gacatcagca atacagacta cttcttcccc
     3061 agatagggat ccactagtcc agtgtggtgg aattctgcag atatccagca cagtggcggc
     3121 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3181 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3241 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3301 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3361 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3421 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3481 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3541 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3601 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3661 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3721 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3781 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3841 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3901 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3961 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4081 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4141 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4201 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4261 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4321 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4381 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4441 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4501 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4561 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4621 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4681 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4741 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4801 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4861 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4921 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4981 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5041 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5101 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5161 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5221 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5281 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5341 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5401 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5461 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5521 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5581 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5641 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5701 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5761 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5821 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5881 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5941 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6001 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6061 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6121 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6181 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6241 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6301 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt
     6361 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6421 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6481 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6541 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6601 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6661 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6721 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6781 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6841 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6901 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6961 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7021 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7081 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7141 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7201 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7261 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7321 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7381 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7441 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7501 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7561 gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
