LOCUS       V023459                 8049 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023459
VERSION     V023459
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..2723
                     /label="CHML(NM_001381853)"
                     /note="CHML(NM_001381853)"
                     /gene="CHML"
     CDS             2736..2801
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2825..3413
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3479..3527
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3729..4157
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4171..4500
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4567..5361
                     /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    5537..5658
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5707..5723)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5731..5747
                     /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(5755..5785)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5800..5821
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6109..6697)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6868..7728)
                     /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(7729..7833)
                     /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 ctcggatccg ccaccatggc ggacaatctt cccacagagt
      781 ttgatgtggt tataataggg acaggtttgc ccgaatccat ccttgcagct gcatgttcaa
      841 gaagtggtca gagggttctg catattgatt caagaagcta ctatggagga aactgggcta
      901 gtttcagctt ttcaggattg ctatcctggt tgaaggagta tcagcaaaac aatgacattg
      961 gggaagaaag tactgttgta tggcaggacc tgatccatga aacagaagaa gccatcactc
     1021 ttcgcaagaa ggatgaaact attcaacaca cagaagcttt ttgctacgcc agtcaggata
     1081 tggaggacaa cgttgaagag attggtgctc tgcagaaaaa tccttctttg ggggtgtcta
     1141 ataccttcac tgaagttctg gattctgcat tacctgaaga aagccagtta tcgtatttta
     1201 atagcgacga aatgcctgca aaacacactc agaaaagtga tacagagatt tcactagaag
     1261 taactgatgt agaggaatca gtggagaagg aaaagtattg tggagataaa acttgtatgc
     1321 acacagtttc agataaagat ggagataaag atgaaagcaa atctacagta gaagataagg
     1381 ccgatgaacc aattagaaat aggattactt actctcaaat agttaaagaa ggcaggaggt
     1441 ttaatattga tttggtgtca aaactgctgt attctcaagg attgctaatt gatcttttaa
     1501 tcaaatcaga tgttagtcgt tatgtagaat ttaaaaatgt cactaggatt cttgcatttc
     1561 gggaaggaaa ggtagaacaa gttccttgtt ccagagcaga tgtctttaat agcaaggaac
     1621 tcaccatggt tgaaaagagg atgctaatga aatttctcac attttgttta gagtatgaac
     1681 aacatcctga tgaataccaa gctttcaggc agtgttcatt ttcagaatac ttaaaaacta
     1741 aaaaactaac tcccaacctt caacattttg tactgcactc aattgcaatg acatcagaat
     1801 catcttgcac tacaatagat ggtcttaacg caactaaaaa cttccttcag tgtctcggac
     1861 ggtttggcaa cacccccttt ttatttccct tgtatggcca aggagaaatt ccccagggtt
     1921 tctgtaggat gtgtgcagtt tttggtggaa tctattgtct tcgtcataaa gtacaatgct
     1981 ttgtagtcga caaagaatct ggacgatgta aagcaattat agatcacttt ggtcaaagaa
     2041 taaatgctaa atattttatt gtggaagaca gttacctttc tgaggaaaca tgctcaaatg
     2101 tgcagtataa gcagatctct agggcagtac tcattacaga tcagtctata ctaaagacag
     2161 atttagatca gcagacttcc attctgatag ttcctccagc agagccagga gcttgtgctg
     2221 tacgggtcac agaattatgt tcttcaacca tgacatgcat gaaggacacc tatctggtac
     2281 atttgacatg ttcatcttct aaaacagcaa gagaagactt agaatcagtg gtgaagaaat
     2341 tattcactcc gtatactgaa acagaaataa acgaggaaga acttacaaag ccaagactct
     2401 tgtgggctct ttattttaat atgagagatt cctcgggaat cagcagaagc tcgtataatg
     2461 gcttgccttc caatgtttat gtctgctctg ggcctgactg tggcctggga aatgagcatg
     2521 ctgtcaagca agctgaaaca cttttccagg agatctttcc aactgaagaa ttctgccctc
     2581 cacctccaaa tccagaagac attatctttg atggtgatga taagcagcca gaggctcctg
     2641 gaaccaataa tgtagtaatg gccaaactag aatcctctga ggaaagcaaa aacctagaaa
     2701 gcccagagaa gcaccttcaa aatctcgagt ctagagacta caaagaccat gacggtgatt
     2761 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     2821 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2881 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2941 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3001 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3061 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3121 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3181 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3241 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3301 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3361 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     3421 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     3481 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     3541 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3601 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3661 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     3721 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3781 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3841 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     3901 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3961 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4021 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4081 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     4141 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4321 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4381 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4441 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4501 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4561 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4621 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4681 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4741 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4801 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4861 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4921 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4981 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5041 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5101 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5161 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5221 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5281 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5341 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     5401 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5461 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5521 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5581 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5641 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5701 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5761 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5821 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5881 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5941 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6001 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6061 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6121 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6181 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6241 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6301 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6361 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6421 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6481 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6541 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6601 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6661 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6721 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6781 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6841 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6901 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6961 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7021 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7081 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7141 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7201 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7261 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7321 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7381 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7441 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7501 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7561 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7621 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7681 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7741 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7801 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7861 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7921 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7981 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     8041 ctgcttagg
//
