LOCUS       V036615                 6400 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036615
VERSION     V036615
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             931..1836
                     /label="ALKBH4(NM_017621)"
                     /note="ALKBH4(NM_017621)"
                     /gene="ALKBH4"
     CDS             1843..1932
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    1967..2191
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2237..2665
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2679..3008
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3075..3869
                     /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    4043..4164
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4213..4229)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4237..4253
                     /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(4261..4291)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4306..4327
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4615..5200)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5371..6231)
                     /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(6232..6336)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggcggcgg ctgccgccga gacccccgaa
      961 gtccttcggg aatgcggttg caagggcatc cggacctgtc tgatctgcga gcggcagcgc
     1021 ggcagtgacc cgccctggga gctgccccca gcgaaaacat accgtttcat ttactgctcc
     1081 gacaccggct gggccgtggg cacagaggag tctgactttg agggctgggc cttccccttc
     1141 ccaggagtga tgctgatcga ggactttgtg acccgggagg aagaagccga gttggtgcgg
     1201 ctcatggacc gtgacccctg gaagctctcc cagtctggac ggaggaagca ggactatggc
     1261 cccaaagtca actttcggaa acagaagcta aagaccgagg gcttctgcgg cctccccagc
     1321 ttcagccggg aggtggtgcg gaggatgggc ctctacccgg ggctggaggg cttccggccc
     1381 gtcgagcagt gcaacctgga ctactgcccc gagcggggct ctgccattga cccccacctg
     1441 gacgacgcct ggctgtgggg ggagcggctg gtcagcctca acctcctgtc ccccaccgtg
     1501 ctgtccatgt gtcgggaggc gcccgggagc ctgctcctct gctcggcccc gtcggctgcc
     1561 ccggaggcct tggtggacag cgtgatagca cccagccggt cggtgctatg ccaggaggtg
     1621 gaggtggcca tccccttacc cgcccgctcc ctgctggtcc tcaccggggc ggcacggcac
     1681 cagtggaagc atgccatcca ccgcagacac atcgaggccc gccgcgtctg cgtcactttc
     1741 cgggagctgt cggctgagtt tggccctgga gggaggcagc aagagctggg ccaggaactg
     1801 ctgcggatcg ccctctcctt ccagggaaga cccgtgctcg aggaacaaaa actaatatca
     1861 gaggaagatt tggaacagaa actaatcagt gaagaagatt tagagcaaaa gttaatttcc
     1921 gaggaggact tatgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     1981 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2041 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2101 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2161 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2221 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2281 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2341 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2401 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2461 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2521 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2581 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2641 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2701 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2761 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2821 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2881 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2941 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3001 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     3061 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3121 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3181 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3241 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3301 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3361 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3421 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3481 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3541 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3601 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3661 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3721 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3781 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3841 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     3901 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     3961 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4021 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4081 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4141 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4201 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4261 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4321 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4381 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4441 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4501 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4561 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4621 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4681 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4741 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4801 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4861 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4921 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4981 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5041 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5101 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     5161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6361 tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
