LOCUS       V024828                 8170 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024828
VERSION     V024828
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..3660
                     /label="ZBTB16(NM_001018011)"
                     /note="ZBTB16(NM_001018011)"
                     /gene="ZBTB16"
     polyA_signal    3737..3961
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4007..4435
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4449..4778
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4845..5639
                     /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    5813..5934
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5983..5999)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6007..6023
                     /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(6031..6061)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6076..6097
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6385..6970)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7141..8001)
                     /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(8002..8106)
                     /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 catggatctg acaaaaatgg gcatgatcca gctgcagaac
     1681 cctagccacc ccacggggct actgtgcaag gccaaccaga tgcggctggc cgggactttg
     1741 tgcgatgtgg tcatcatggt ggacagccag gagttccacg cccaccggac ggtgctggcc
     1801 tgcaccagca agatgtttga gatcctcttc caccgcaata gtcaacacta tactttggac
     1861 ttcctctcgc caaagacctt ccagcagatt ctggagtatg catatacagc cacgctgcaa
     1921 gccaaggcgg aggacctgga tgacctgctg tatgcggccg agatcctgga gatcgagtac
     1981 ctggaggaac agtgcctgaa gatgctggag accatccagg cctcagacga caatgacacg
     2041 gaggccacca tggccgatgg cggggccgag gaagaagagg accgcaaggc tcggtacctc
     2101 aagaacatct tcatctcgaa gcattccagc gaggagagtg ggtatgccag tgtggctgga
     2161 cagagcctcc ctgggcccat ggtggaccag agcccttcag tctccacttc atttggtctt
     2221 tcagccatga gtcccaccaa ggctgcagtg gacagtttga tgaccatagg acagtctctc
     2281 ctgcagggaa ctcttcagcc acctgcaggg cccgaggagc caactctggc tgggggtggg
     2341 cggcaccctg gggtggctga ggtgaagacg gagatgatgc aggtggatga ggtgcccagc
     2401 caggacagcc ctggggcagc cgagtccagc atctcaggag ggatggggga caaggttgag
     2461 gaaagaggca aagaggggcc tgggaccccg actcgaagca gcgtcatcac cagtgctagg
     2521 gagctacact atgggcgaga ggagagtgcc gagcaggtgc cacccccagc tgaggctggc
     2581 caggccccca ctggccgacc tgagcaccca gcacccccgc ctgagaagca tctgggcatc
     2641 tactccgtgt tgcccaacca caaggctgac gctgtattga gcatgccgtc ttccgtgacc
     2701 tctggcctcc acgtgcagcc tgccctggct gtctccatgg acttcagcac ctatgggggg
     2761 ctgctgcccc agggcttcat ccagagggag ctgttcagca agctggggga gctggctgtg
     2821 ggcatgaagt cagagagccg gaccatcgga gagcagtgca gcgtgtgtgg ggtcgagctt
     2881 cctgataacg aggctgtgga gcagcacagg aagctgcaca gtgggatgaa gacgtacggg
     2941 tgcgagctct gcgggaagcg gttcctggat agtttgcggc tgagaatgca cttactggct
     3001 cattcagcgg gtgccaaagc ctttgtctgt gatcagtgcg gtgcacagtt ttcgaaggag
     3061 gatgccctgg agacacacag gcagacccat actggcactg acatggccgt cttctgtctg
     3121 ctgtgtggga agcgcttcca ggcgcagagc gcactgcagc agcacatgga ggtccacgcg
     3181 ggcgtgcgca gctacatctg cagtgagtgc aaccgcacct tccccagcca cacggctctc
     3241 aaacgccacc tgcgctcaca tacaggcgac cacccctacg agtgtgagtt ctgtggcagc
     3301 tgcttccggg atgagagcac actcaagagc cacaaacgca tccacacggg tgagaaaccc
     3361 tacgagtgca atggctgtgg caagaagttc agcctcaagc atcagctgga gacgcactat
     3421 agggtgcaca caggtgagaa gccctttgag tgtaagctct gccaccagcg ctcccgggac
     3481 tactcggcca tgatcaagca cctgagaacg cacaacggcg cctcgcccta ccagtgcacc
     3541 atctgcacag agtactgccc cagcctctcc tccatgcaga agcacatgaa gggccacaag
     3601 cccgaggaga tcccgcccga ctggaggata gagaagacgt acctctacct gtgctatgtg
     3661 tgaggatcct gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3721 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3781 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3841 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3901 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3961 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4021 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4081 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4141 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4201 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4261 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4321 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4381 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4441 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4501 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4561 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4621 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4681 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4741 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4801 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4861 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4921 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4981 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5041 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5101 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5161 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5221 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5281 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5341 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5401 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5461 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5521 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5581 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5641 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5701 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5761 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5821 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5881 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5941 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6001 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6061 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6121 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6181 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6241 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6301 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6901 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6961 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7021 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7081 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7141 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7201 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7261 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7321 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7381 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7441 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7501 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7561 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7621 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7681 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7741 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7801 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7861 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7921 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7981 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8101 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     8161 cccctatggt
//
