LOCUS       V045644                 7077 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045644
VERSION     V045644
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1757
                     /label="Mogat2(NM_177448)"
                     /note="Mogat2(NM_177448)"
                     /gene="Mogat2"
     CDS             1764..1829
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1853..2441
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2507..2555
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2757..3185
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3199..3528
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3595..4389
                     /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    4565..4686
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4735..4751)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4759..4775
                     /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(4783..4813)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4828..4849
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5137..5725)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5896..6756)
                     /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(6757..6861)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggt ggagttcgcc cccctgttgg
      781 taccatggga gcgcaggtta cagaccttcg cggtccttca gtgggtcttc tccttcctgg
      841 ccttggccca gctctgcatc gtcatcttcg taggcctcct attcacaagg ttctggctct
      901 tctctgtcct gtatgccacc tggtggtacc tggactggga caagccgcgg cagggaggcc
      961 ggcccatcca gttcttcaga cgcttggcca tatggaagta catgaaggat tatttccctg
     1021 tctctttggt caagacagct gagctggacc cttcccggaa ctacatcgcg ggcttccacc
     1081 cccatggagt cctagcagct ggagcctttc ttaacctgtg cactgaaagc acgggcttta
     1141 cctcgctttt cccgggcatc cgctcctatc tgatgatgct gactgtgtgg ttccgggccc
     1201 ccttcttccg agattacatc atgtctgggg ggctggtctc atcagaaaag gtgagtgccg
     1261 atcacattct gtccaggaag ggcggcggga acttgcttgc catcatcgtt gggggcgcgc
     1321 aggaggcact ggacgccagg cctggagcct acaggctgct gctgaagaat cgcaagggct
     1381 tcatcaggct cgccctgatg catggggcag ctcttgtgcc aatcttctcc tttggagaaa
     1441 acaacctgtt caaccaggtt gagaacaccc ctggtacctg gctgcgctgg atccagaacc
     1501 ggctacagaa gatcatgggc atctccctcc ctctcttcca cggcagaggt gtcttccagt
     1561 acagctttgg cctcatgccc ttccgccagc ccatcaccac catagtgggg aagcccatcg
     1621 aggtgcagat gacaccacag ccctcaaggg aggaggtgga ccggcttcat cagcgctata
     1681 tcaaggagct ctgcaagctc tttgaggagc acaaactcaa gttcaacgtc cctgaggacc
     1741 agcatctgga gttctgcgga tccgactaca aagaccatga cggtgattat aaagatcatg
     1801 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     1861 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1921 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1981 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2041 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2101 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2161 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2221 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     2281 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2341 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2401 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     2461 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     2521 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     2581 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2641 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2701 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     2761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2821 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2881 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     2941 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3061 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3121 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3181 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3301 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3361 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3421 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3481 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3541 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3601 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3661 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3721 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3781 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3841 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3901 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3961 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4021 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4081 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4141 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4201 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4261 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4321 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4381 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4441 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4501 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4561 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4621 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4681 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4741 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4801 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4861 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4921 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4981 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5041 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5101 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5161 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5221 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5281 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     5341 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5401 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5461 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5521 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5581 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5641 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5701 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5761 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5821 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5881 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5941 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6001 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6061 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6121 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6181 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6241 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6301 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6361 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6421 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6481 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6541 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6601 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6661 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6721 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6781 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6841 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6901 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     6961 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     7021 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
