LOCUS       V021419                 7649 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021419
VERSION     V021419
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             647..2329
                     /label="Ces2c(NM_145603)"
                     /note="Ces2c(NM_145603)"
                     /gene="Ces2c"
     CDS             2336..2401
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2425..3013
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3079..3127
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3329..3757
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3771..4100
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4167..4961
                     /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    5137..5258
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5307..5323)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5331..5347
                     /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(5355..5385)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5400..5421
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5709..6297)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6468..7328)
                     /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(7329..7433)
                     /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 gctctggcta gccaccatga cacggaacca
      661 actacataac tggctgaatg ctgggttctt cgggctgctg cttcttctta tccatgtgca
      721 gggtcaggac tcaccagagg ccaaccccat cagaaacaca catacaggac agatccaagg
      781 cagccttatc cacgtgaagg acactaaagc tggtgtccac accttcctgg gaatcccctt
      841 tgccaaacct cctgtaggac cactgcgatt tgcacctcct gaggcccctg aaccatggag
      901 tggtgtgaga gatggaactg cacatccggc catgtgtctg caaaatcttg acatgctgaa
      961 tgaagcaggc ctgccagata tgaaaatgat gctgtcttcc ttccctatgt ctgaggactg
     1021 cctgtatctc aacatctaca ccccagccca tgcccatgag ggctctaacc tgcctgtgat
     1081 ggtgtggatc catggaggtg cactggttat aggaatggct tccatgtttg atggatctct
     1141 attgacagtc aatgaggact tggtggttgt tactatccag tatcgtctgg gtgtcctggg
     1201 ctttttcagc actggagacc agcatgccag aggcaactgg ggatacctgg accaagcagc
     1261 tgccctacgc tgggtccagc agaacatcgc tcactttgga ggcaaccctg accgggtcac
     1321 tatttttggc gagtctgcag gtggcacaag tgtgtcttca catgttgtgt cccccatgtc
     1381 ccaagggctc ttccatggtg ccatcatgga gagtggggtg gccctgcttc ctgaccttat
     1441 ctctgaaacc tctgagatgg tttccactac ggtggccaag ctttctggat gtgaggctat
     1501 ggattcccag gccctggtgc gctgtctgag aggcaaaagt gaagcagaga ttctggctat
     1561 taacaaggtc ttcaagatga tccctgctgt ggtggatggg gagttcttcc ccaggcatcc
     1621 caaagagttg ttggcatctg aggattttca ccctgtcccc agcatcattg gtgtcaacaa
     1681 tgatgagttt ggctggtcca ttcctgtggt catgggctct gctcagatga taaaaggaat
     1741 aaccagagag aacctgcagg ctgttctgaa ggatacagca gtacaaatga tgctgcctcc
     1801 tgagtgtagt gacctgctaa tggaagagta catgggggac actgaggatg cccagaccct
     1861 ccaaatacag ttcacagaga tgatgggaga tttcatgttt gtgatccctg cactccaagt
     1921 agcacatttt caacgctccc atgcccctgt ctacttctat gagttccaac atccacccag
     1981 ctacttcaag gatgtcagac caccccacgt gaaggctgac catgctgatg agattccttt
     2041 tgtctttgcg tccttcttct ggggcatgaa acttgacttc actgaggagg aggagctgct
     2101 gagcaggagg atgatgaagt actgggccaa ctttgcacga catgggaacc ccaacagtga
     2161 gggtctaccc tactggcctg tgatggacca tgatgagcag tacctgcaac tggacatcca
     2221 gcctgctgtg ggtcgagccc tgaaggctgg aaggctgcag ttctggacca agactctgcc
     2281 ccagaagatc caggagctaa aggcttctca ggacaagcac agggagcttc tcgaggacta
     2341 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     2401 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     2461 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2521 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2581 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2641 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2701 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2761 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     2821 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2881 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2941 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3001 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     3061 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     3121 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3181 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3241 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     3301 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3361 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3421 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3481 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3541 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3601 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3661 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     3721 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3781 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3841 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3901 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3961 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4021 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4081 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4141 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4201 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4261 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4321 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4381 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4441 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4501 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4561 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4621 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4681 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4741 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4801 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4861 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4921 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     4981 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5041 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5101 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5161 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5221 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5281 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5341 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5401 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5461 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5521 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5581 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5641 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5701 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5761 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5821 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5881 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     5941 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6001 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6061 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6121 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6181 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6241 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6301 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6361 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6421 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6481 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6541 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6601 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6661 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6721 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6781 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6841 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6901 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6961 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7021 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7081 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7141 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7201 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7261 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7321 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7381 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7441 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     7501 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     7561 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     7621 ccgacaattg catgaagaat ctgcttagg
//
