LOCUS       V037036                 9791 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037036
VERSION     V037036
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"
     CDS             775..4467
                     /label="CFH(NM_000186)"
                     /note="CFH(NM_000186)"
                     /gene="CFH"
     CDS             4474..4497
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     misc_feature    4567..5155
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5221..5269
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5471..5899
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5913..6242
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6309..7103
                     /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    7279..7400
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7449..7465)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7473..7489
                     /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(7497..7527)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7542..7563
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7851..8439)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8610..9470)
                     /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(9471..9575)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatgaga
      781 cttctagcaa agattatttg ccttatgtta tgggctattt gtgtagcaga agattgcaat
      841 gaacttcctc caagaagaaa tacagaaatt ctgacaggtt cctggtctga ccaaacatat
      901 ccagaaggca cccaggctat ctataaatgc cgccctggat atagatctct tggaaatgta
      961 ataatggtat gcaggaaggg agaatgggtt gctcttaatc cattaaggaa atgtcagaaa
     1021 aggccctgtg gacatcctgg agatactcct tttggtactt ttacccttac aggaggaaat
     1081 gtgtttgaat atggtgtaaa agctgtgtat acatgtaatg aggggtatca attgctaggt
     1141 gagattaatt accgtgaatg tgacacagat ggatggacca atgatattcc tatatgtgaa
     1201 gttgtgaagt gtttaccagt gacagcacca gagaatggaa aaattgtcag tagtgcaatg
     1261 gaaccagatc gggaatacca ttttggacaa gcagtacggt ttgtatgtaa ctcaggctac
     1321 aagattgaag gagatgaaga aatgcattgt tcagacgatg gtttttggag taaagagaaa
     1381 ccaaagtgtg tggaaatttc atgcaaatcc ccagatgtta taaatggatc tcctatatct
     1441 cagaagatta tttataagga gaatgaacga tttcaatata aatgtaacat gggttatgaa
     1501 tacagtgaaa gaggagatgc tgtatgcact gaatctggat ggcgtccgtt gccttcatgt
     1561 gaagaaaaat catgtgataa tccttatatt ccaaatggtg actactcacc tttaaggatt
     1621 aaacacagaa ctggagatga aatcacgtac cagtgtagaa atggttttta tcctgcaacc
     1681 cggggaaata cagcaaaatg cacaagtact ggctggatac ctgctccgag atgtaccttg
     1741 aaaccttgtg attatccaga cattaaacat ggaggtctat atcatgagaa tatgcgtaga
     1801 ccatactttc cagtagctgt aggaaaatat tactcctatt actgtgatga acattttgag
     1861 actccgtcag gaagttactg ggatcacatt cattgcacac aagatggatg gtcgccagca
     1921 gtaccatgcc tcagaaaatg ttattttcct tatttggaaa atggatataa tcaaaatcat
     1981 ggaagaaagt ttgtacaggg taaatctata gacgttgcct gccatcctgg ctacgctctt
     2041 ccaaaagcgc agaccacagt tacatgtatg gagaatggct ggtctcctac tcccagatgc
     2101 atccgtgtca aaacatgttc caaatcaagt atagatattg agaatgggtt tatttctgaa
     2161 tctcagtata catatgcctt aaaagaaaaa gcgaaatatc aatgcaaact aggatatgta
     2221 acagcagatg gtgaaacatc aggatcaatt acatgtggga aagatggatg gtcagctcaa
     2281 cccacgtgca ttaaatcttg tgatatccca gtatttatga atgccagaac taaaaatgac
     2341 ttcacatggt ttaagctgaa tgacacattg gactatgaat gccatgatgg ttatgaaagc
     2401 aatactggaa gcaccactgg ttccatagtg tgtggttaca atggttggtc tgatttaccc
     2461 atatgttatg aaagagaatg cgaacttcct aaaatagatg tacacttagt tcctgatcgc
     2521 aagaaagacc agtataaagt tggagaggtg ttgaaattct cctgcaaacc aggatttaca
     2581 atagttggac ctaattccgt tcagtgctac cactttggat tgtctcctga cctcccaata
     2641 tgtaaagagc aagtacaatc atgtggtcca cctcctgaac tcctcaatgg gaatgttaag
     2701 gaaaaaacga aagaagaata tggacacagt gaagtggtgg aatattattg caatcctaga
     2761 tttctaatga agggacctaa taaaattcaa tgtgttgatg gagagtggac aactttacca
     2821 gtgtgtattg tggaggagag tacctgtgga gatatacctg aacttgaaca tggctgggcc
     2881 cagctttctt cccctcctta ttactatgga gattcagtgg aattcaattg ctcagaatca
     2941 tttacaatga ttggacacag atcaattacg tgtattcatg gagtatggac ccaacttccc
     3001 cagtgtgtgg caatagataa acttaagaag tgcaaatcat caaatttaat tatacttgag
     3061 gaacatttaa aaaacaagaa ggaattcgat cataattcta acataaggta cagatgtaga
     3121 ggaaaagaag gatggataca cacagtctgc ataaatggaa gatgggatcc agaagtgaac
     3181 tgctcaatgg cacaaataca attatgccca cctccacctc agattcccaa ttctcacaat
     3241 atgacaacca cactgaatta tcgggatgga gaaaaagtat ctgttctttg ccaagaaaat
     3301 tatctaattc aggaaggaga agaaattaca tgcaaagatg gaagatggca gtcaatacca
     3361 ctctgtgttg aaaaaattcc atgttcacaa ccacctcaga tagaacacgg aaccattaat
     3421 tcatccaggt cttcacaaga aagttatgca catgggacta aattgagtta tacttgtgag
     3481 ggtggtttca ggatatctga agaaaatgaa acaacatgct acatgggaaa atggagttct
     3541 ccacctcagt gtgaaggcct tccttgtaaa tctccacctg agatttctca tggtgttgta
     3601 gctcacatgt cagacagtta tcagtatgga gaagaagtta cgtacaaatg ttttgaaggt
     3661 tttggaattg atgggcctgc aattgcaaaa tgcttaggag aaaaatggtc tcaccctcca
     3721 tcatgcataa aaacagattg tctcagttta cctagctttg aaaatgccat acccatggga
     3781 gagaagaagg atgtgtataa ggcgggtgag caagtgactt acacttgtgc aacatattac
     3841 aaaatggatg gagccagtaa tgtaacatgc attaatagca gatggacagg aaggccaaca
     3901 tgcagagaca cctcctgtgt gaatccgccc acagtacaaa atgcttatat agtgtcgaga
     3961 cagatgagta aatatccatc tggtgagaga gtacgttatc aatgtaggag cccttatgaa
     4021 atgtttgggg atgaagaagt gatgtgttta aatggaaact ggacggaacc acctcaatgc
     4081 aaagattcta caggaaaatg tgggccccct ccacctattg acaatgggga cattacttca
     4141 ttcccgttgt cagtatatgc tccagcttca tcagttgagt accaatgcca gaacttgtat
     4201 caacttgagg gtaacaagcg aataacatgt agaaatggac aatggtcaga accaccaaaa
     4261 tgcttacatc cgtgtgtaat atcccgagaa attatggaaa attataacat agcattaagg
     4321 tggacagcca aacagaagct ttattcgaga acaggtgaat cagttgaatt tgtgtgtaaa
     4381 cggggatatc gtctttcatc acgttctcac acattgcgaa caacatgttg ggatgggaaa
     4441 ctggagtatc caacttgtgc aaaaagagaa ttcgactaca aggacgacga tgacaagtga
     4501 ctcgagtcta gagggcccct taagggttcg atccctaccg gttagtaatg agtttgatat
     4561 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4621 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4681 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4741 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4801 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4861 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4921 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     4981 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     5041 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     5101 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     5161 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5221 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     5281 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5341 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5401 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     5461 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5521 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5581 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     5641 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5701 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5761 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5821 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     5881 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5941 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     6001 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6061 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     6121 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6181 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     6241 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     6301 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     6361 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     6421 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     6481 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     6541 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     6601 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     6661 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     6721 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     6781 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     6841 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     6901 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     6961 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     7021 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     7081 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     7141 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     7201 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     7261 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     7321 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     7381 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     7441 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     7501 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     7561 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     7621 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7681 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7741 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7801 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7861 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7921 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7981 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     8041 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8101 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8161 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     8221 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     8281 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     8341 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     8401 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     8461 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     8521 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     8581 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     8641 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     8701 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8761 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     8821 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8881 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8941 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     9001 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9061 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9121 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9181 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     9241 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     9301 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     9361 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     9421 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     9481 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     9541 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     9601 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     9661 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     9721 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     9781 atctgcttag g
//
