LOCUS       V018365                 9419 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018365
VERSION     V018365
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             671..4090
                     /label="DDB1(NM_001923)"
                     /note="DDB1(NM_001923)"
                     /gene="DDB1"
     CDS             4106..4171
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4195..4783
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4849..4897
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5099..5527
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5541..5870
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5937..6731
                     /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    6907..7028
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7077..7093)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7101..7117
                     /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(7125..7155)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7170..7191
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7479..8067)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8238..9098)
                     /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(9099..9203)
                     /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 gcgtttaaac ttaagcttgg
      661 taccgccacc atgtcgtaca actacgtggt aacggcccag aagcccaccg ccgtgaacgg
      721 ctgcgtgacc ggacacttta cttcggccga agacttaaac ctgttgattg ccaaaaacac
      781 gagattagag atctatgtgg tcaccgccga ggggcttcgg cccgtcaaag aggtgggcat
      841 gtatgggaag attgcggtca tggagctttt caggcccaag ggggagagca aggacctgct
      901 gtttatcttg acagcgaagt acaatgcctg catcctggag tataaacaga gtggcgagag
      961 cattgacatc attacgcgag cccatggcaa tgtccaggac cgcattggcc gcccctcaga
     1021 gaccggcatt attggcatca ttgaccctga gtgccggatg attggcctgc gtctctatga
     1081 tggccttttc aaggttattc cactagatcg cgataataaa gaactcaagg ccttcaacat
     1141 ccgcctggag gagctgcatg tcattgatgt caagttccta tatggttgcc aagcacctac
     1201 tatttgcttt gtctaccagg accctcaggg gcggcacgta aaaacctatg aggtgtctct
     1261 ccgagaaaag gaattcaata agggcccttg gaaacaggaa aatgtcgaag ctgaagcttc
     1321 catggtgatc gcagtcccag agccctttgg gggggccatc atcattggac aggagtcaat
     1381 cacctatcac aatggtgaca aatacctggc tattgcccct cctatcatca agcaaagcac
     1441 gattgtgtgc cacaatcgag tggaccctaa tggctcaaga tacctgctgg gagacatgga
     1501 aggccggctc ttcatgctgc ttttggagaa ggaggaacag atggatggca ccgtcactct
     1561 caaggatctc cgtgtagaac tccttggaga gacctctatt gctgagtgct tgacatacct
     1621 tgataatggt gttgtgtttg tcgggtctcg cctgggtgac tcccagcttg tgaagctcaa
     1681 cgttgacagt aatgaacaag gctcctatgt agtggccatg gaaaccttta ccaacttagg
     1741 acccattgtc gatatgtgcg tggtggacct ggagaggcag gggcaggggc agctggtcac
     1801 ttgctctggg gctttcaagg aaggttcttt gcggatcatc cggaatggaa ttggaatcca
     1861 cgagcatgcc agcattgact taccaggcat caaaggatta tggccactgc ggtctgaccc
     1921 taatcgtgag actgatgaca ctttggtgct ctcttttgtg ggccagacaa gagttctcat
     1981 gttaaatgga gaggaggtag aagaaaccga actgatgggt ttcgtggatg atcagcagac
     2041 tttcttctgt ggcaacgtgg ctcatcagca gcttatccag atcacttcag catcggtgag
     2101 gttggtctct caagaaccca aagctctggt cagtgaatgg aaggagcctc aggccaagaa
     2161 catcagtgtg gcctcctgca atagcagcca ggtggtggtg gctgtaggca gggccctcta
     2221 ctatctgcag atccatcctc aggagctccg gcagatcagc cacacagaga tggaacatga
     2281 agtggcttgc ttggacatca ccccattagg agacagcaat ggactgtccc ctctttgtgc
     2341 cattggcctc tggacggaca tctcggctcg tatcttgaag ttgccctctt ttgaactact
     2401 gcacaaggag atgctgggtg gagagatcat tcctcgctcc atcctgatga ccacctttga
     2461 gagtagccat tacctccttt gtgccttggg agatggagcg cttttctact ttgggctcaa
     2521 cattgagaca ggtctgttga gcgaccgtaa gaaggtgact ttgggcaccc agcccaccgt
     2581 attgaggact tttcgttctc tttctaccac caacgtcttt gcttgttctg accgccccac
     2641 tgtcatctat agcagcaacc acaaattggt cttctcaaat gtcaacctca aggaagtgaa
     2701 ctacatgtgt cccctcaatt cagatggcta tcctgacagc ctggcgctgg ccaacaatag
     2761 caccctcacc attggcacca tcgatgagat ccagaagctg cacattcgca cagttcccct
     2821 ctatgagtct ccaaggaaga tctgctacca ggaagtgtcc cagtgtttcg gggtcctctc
     2881 cagccgcatt gaagtccaag acacgagtgg gggcacgaca gccttgaggc ccagcgctag
     2941 cacccaggct ctgtccagca gtgtaagctc cagcaagctg ttctccagca gcactgctcc
     3001 tcatgagacc tcctttggag aagaggtgga ggtgcacaac ctacttatca ttgaccaaca
     3061 cacctttgaa gtgcttcatg cccaccagtt tctgcagaat gaatatgccc tcagtctggt
     3121 ttcctgcaag ctgggcaaag accccaacac ttacttcatt gtgggcacag caatggtgta
     3181 tcctgaagag gcagagccca agcagggtcg cattgtggtc tttcagtatt cggatggaaa
     3241 actacagact gtggctgaaa aggaagtgaa aggggccgtg tactctatgg tggaatttaa
     3301 cgggaagctg ttagccagca tcaatagcac ggtgcggctc tatgagtgga caacagagaa
     3361 ggagctgcgc actgagtgca accactacaa caacatcatg gccctctacc tgaagaccaa
     3421 gggcgacttc atcctggtgg gcgaccttat gcgctcagtg ctgctgcttg cctacaagcc
     3481 catggaagga aactttgaag agattgctcg agactttaat cccaactgga tgagtgctgt
     3541 ggaaatcttg gatgatgaca attttctggg ggctgaaaat gcctttaact tgtttgtgtg
     3601 tcaaaaggat agcgctgcca ccactgacga ggagcggcag cacctccagg aggttggtct
     3661 tttccacctg ggcgagtttg tcaatgtctt ttgccacggc tctctggtaa tgcagaatct
     3721 gggtgagact tccaccccca cacaaggctc ggtgctcttc ggcacggtca acggcatgat
     3781 agggctggtg acctcactgt cagagagctg gtacaacctc ctgctggaca tgcagaatcg
     3841 actcaataaa gtcatcaaaa gtgtggggaa gatcgagcac tccttctgga gatcctttca
     3901 caccgagcgg aagacagaac cagccacagg tttcatcgac ggtgacttga ttgagagttt
     3961 cctggatatt agccgcccca agatgcagga ggtggtggca aacctacagt atgacgatgg
     4021 cagcggtatg aagcgagagg ccactgcaga cgacctcatc aaggttgtgg aggagctaac
     4081 tcggatccat cagcggccgc tcgaggacta caaagaccat gacggtgatt ataaagatca
     4141 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     4201 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4261 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4321 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4381 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4441 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4501 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4561 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     4621 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4681 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4741 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     4801 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4861 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4921 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4981 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5041 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     5101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5161 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5221 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     5281 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5401 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5461 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5521 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5581 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5641 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5701 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5761 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5821 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5881 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5941 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     6001 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     6061 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     6121 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     6181 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     6241 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6301 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6361 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6421 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6481 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6541 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6601 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6661 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6721 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     6781 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6841 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6901 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6961 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     7021 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     7081 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7141 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7201 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7261 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7321 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7381 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7441 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     7681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9121 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9181 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     9241 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     9301 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     9361 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
