LOCUS       V040112                 8140 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040112
VERSION     V040112
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..3630
                     /label="ALOX12(NM_000697)"
                     /note="ALOX12(NM_000697)"
                     /gene="ALOX12"
     polyA_signal    3707..3931
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3977..4405
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4419..4748
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4815..5609
                     /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    5783..5904
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5953..5969)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5977..5993
                     /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(6001..6031)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6046..6067
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6355..6940)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7111..7971)
                     /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(7972..8076)
                     /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 catgggccgc taccgcatcc gcgtggccac cggggcctgg
     1681 ctcttctccg ggtcgtacaa ccgcgtgcag ctttggctgg tcgggacgcg cggggaggcg
     1741 gagctggagc tgcagctgcg gcccgcgcgg ggcgaggagg aggagtttga tcatgacgtt
     1801 gcagaggact tggggctcct gcagttcgtg aggctgcgca agcaccactg gctggtggac
     1861 gacgcgtggt tctgcgaccg catcacggtg cagggccctg gagcctgcgc ggaggtggcc
     1921 ttcccgtgct accgctgggt gcagggcgag gacatcctga gcctgcccga gggcaccgcc
     1981 cgcctgccag gagacaatgc tttggacatg ttccagaagc atcgagagaa ggaactgaaa
     2041 gacagacagc agatctactg ctgggccacc tggaaggaag ggttacccct gaccatcgct
     2101 gcagaccgta aggatgatct acctccaaat atgagattcc atgaggagaa gaggctggac
     2161 tttgaatgga cactgaaggc aggggctctg gagatggccc tcaaacgtgt ttacaccctc
     2221 ctgagctcct ggaactgcct agaagacttt gatcagatct tctggggcca gaagagtgcc
     2281 ctggctgaga aggttcgcca gtgctggcag gatgatgagt tgttcagcta ccagttcctc
     2341 aatggtgcca accccatgct gttgagacgc tcgacctctc tgccctccag gctagtgctg
     2401 ccctcgggga tggaagagct tcaggctcaa ctggagaaag aacttcagaa tggttccctg
     2461 tttgaagctg acttcatcct tctggatgga attccagcca acgtgatccg aggagagaag
     2521 caatacctgg ctgcccccct cgttatgctg aagatggagc ccaatgggaa gctgcagccc
     2581 atggtcatcc agattcagcc tcccaacccc agctctccaa ccccaacact gttcctgccc
     2641 tcagaccccc cacttgcctg gctcctggca aagtcctggg tccgaaattc agatttccaa
     2701 ctgcacgaga tccagtatca cttgctgaac actcacctgg tggctgaggt catcgctgtc
     2761 gccaccatgc ggtgcctccc aggactgcac cccatcttca agttcctgat cccccatatc
     2821 cgctacacca tggaaatcaa cacccgggcc cggacccaac tcatctcaga tggaggaatt
     2881 tttgataagg cagtgagcac aggtggaggg ggccatgtac agttgctccg tcgggcggca
     2941 gctcagctga cctactgctc cctctgtcct cctgacgacc tggctgaccg gggcctgctg
     3001 ggactcccag gtgctctcta tgcccatgat gctttacggc tctgggagat cattgccagg
     3061 tatgtggagg ggatcgtcca cctcttctac caaagggatg acatagtgaa gggggaccct
     3121 gagctgcagg cctggtgtcg ggagatcacg gaggtggggc tgtgccaggc ccaggaccga
     3181 ggtttccctg tctccttcca gtcccagagt caactctgcc atttcctcac catgtgcgtc
     3241 ttcacgtgca ctgcccagca tgccgccatc aaccagggcc agctggactg gtatgcctgg
     3301 gtccctaatg ctccatgcac aatgcggatg cccccaccca ccaccaagga agatgtgacg
     3361 atggccacag tgatggggtc actacctgat gtccggcagg cctgtcttca aatggccatc
     3421 tcatggcatc tgagtcgccg ccagccagac atggtgcctc tggggcacca caaagaaaaa
     3481 tatttctcag gccccaagcc caaagctgtg ctaaaccaat tccgaacaga tttggaaaag
     3541 ctggaaaagg agattacagc ccggaatgag caacttgact ggccctatga atatctgaag
     3601 cccagctgca tagagaacag tgtcaccatc tgaggatcct gatatccagc acagtggcgg
     3661 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3721 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3781 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3841 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3901 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3961 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4021 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4081 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4141 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4201 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4261 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4321 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4381 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4501 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4561 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4621 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4681 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4741 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4801 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4861 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4921 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4981 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5041 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5101 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5161 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5221 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5281 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5341 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5401 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5461 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5521 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5581 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5641 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5701 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5761 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5821 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5881 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5941 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6001 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6061 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6121 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6181 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6241 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6301 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6361 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6421 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6481 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6541 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6601 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6661 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6721 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6781 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6841 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6901 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6961 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7021 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7081 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7141 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7201 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7261 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7321 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7381 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7441 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7501 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7561 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7621 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7681 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7741 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7801 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7861 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7921 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7981 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8041 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8101 tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
