LOCUS       V044602                 8929 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044602
VERSION     V044602
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..4419
                     /label="Ddb1(NM_001410219)"
                     /note="Ddb1(NM_001410219)"
                     /gene="Ddb1"
     CDS             4426..4491
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4529..4753
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4799..5227
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5241..5570
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5637..6431
                     /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    6605..6726
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6775..6791)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6799..6815
                     /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(6823..6853)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6868..6889
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7177..7762)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7933..8793)
                     /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(8794..8898)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgtcgt acaactacgt cgtaacggcg cagaagccta ctgcagtgaa cggctgtgtg
     1021 accggacact ttacttcagc ggaagatctg aacctgttga ttgctaaaaa cacaagatta
     1081 gagatctatg tggtcactgc ggagggtctt cggccagtca aagaggtggg aatgtacggg
     1141 aagattgcag tcatggagct cttcaggccc aagggggaga gcaaagacct gctgttcatc
     1201 ttgactgcta aatacaatgc ctgtatcttg gagtataagc agagtggcga aagcattgac
     1261 atcattacaa gagcccatgg caatgtccag gaccgcattg gtcgcccctc ggagactggc
     1321 atcattggca tcattgaccc tgagtgccgt atgattggcc ttcgactcta cgatggcctt
     1381 ttcaaggtca ttccactaga ccgagacaat aaagagctca aggcctttaa catccgccta
     1441 gaggaactgc acgtcattga tgtcaagttc ctgtatggtt gtcaagcacc taccatttgc
     1501 tttgtctacc agcctcttaa aggtgacttt ggcagcccca agactgatga tcctcagggg
     1561 cggcatgtga aaacctatga ggtgtctctt cgagagaagg agttcaacaa gggcccttgg
     1621 aaacaggaga atgtagaagc cgaagcttca atggtgatag cagtcccaga gccttttgga
     1681 ggtgccatca tcattggaca ggaatcaatc acctatcata acggtgataa atacctggca
     1741 attgctcctc ctatcattaa gcaaagcact attgtgtgcc ataaccgggt ggatcctaat
     1801 ggctcgcggt accttctggg agatatggag ggacggcttt tcatgctgct cttagagaag
     1861 gaggagcaga tggatggcac tgtcaccctc aaggaccttc gagtggaact cctcggagag
     1921 acctccattg ctgagtgcct aacttatctt gataatggtg ttgtatttgt cgggtctcgc
     1981 ctaggtgact cccagctcgt aaagctcaat gtcgacagca acgaacaagg ctcctatgta
     2041 gtggccatgg aaacctttac caatttagga cccattgtgg atatgtgtgt ggtggacctg
     2101 gagaggcagg gacagggtca gctggtcact tgctctggag ctttcaagga aggttccttg
     2161 cggatcatcc ggaacggcat tggaatccat gagcatgcca gcattgactt accaggcatc
     2221 aaaggtttat ggcctctgcg ctctgacccc ggtcgggaga cggatgacac tttggtgctc
     2281 tcttttgtgg gccagacaag agttctcatg ctaaacggag aggaagtgga agagacagaa
     2341 ctgatgggct ttgtggacga tcagcagact ttcttttgtg gcaacgtggc tcatcagcaa
     2401 cttatccaga tcacctcagc atctgtgagg ttggtgtctc aagagcccaa agctttggtc
     2461 agcgagtgga aagagcctca gggcaagaac atcagtgtgg cctcctgtaa cagcagccag
     2521 gtggttgttg ccgtgggaag ggcactatat taccttcaga tccaccctca ggagctccgg
     2581 caaatcagcc acacagagat ggaacatgaa gtggcatgct tggatatcac cccattaggg
     2641 gacagcaatg gcctctcccc actttgtgcc attggacttt ggacagacat ctcagctcgt
     2701 atcctgaagc tcccatcctt tgaactacta cacaaggaga tgctgggtgg agagatcatt
     2761 cctcgatcca tcctgatgac cacttttgag agtagccact acctcctttg tgccttggga
     2821 gatggggctc ttttctactt tgggctcaac atcgagacag gtttactgag tgaccgtaaa
     2881 aaggtgactt tgggcaccca gcccacggtg ttgaggacat tccgttctct ttctaccacc
     2941 aacgtctttg cttgctctga ccggcccact gtcatctata gcagcaatca caagttggtc
     3001 ttctccaatg tcaacctcaa ggaggtgaac tatatgtgtc ctctcaactc agatggctat
     3061 cctgacagtc tggcattggc caataacagc actctcacca ttggcaccat cgatgagatc
     3121 cagaagctcc atattcgcac agttcccctc tatgagtctc ccaggaagat ctgctatcag
     3181 gaagtgtctc agtgctttgg ggtcctttcc agccgcattg aagtccaaga tagcagtgga
     3241 ggcactactg ctttgaggcc cagtgccagc acccaggccc tgtccagcag cgtcagctcc
     3301 agcaaacttt tctccagcag tacggctcct cacgagacct cctttggaga agaagtggag
     3361 gtgcacaacc tcctcatcat tgaccagcac acctttgaag tgcttcacgc ccatcagttt
     3421 ctacaaaacg agtatgccct cagcctggtc tcctgcaagt tgggtaaaga ccccaacact
     3481 tacttcattg tgggcacagc catggtatat cctgaagagg cagagcccaa gcagggtcgt
     3541 attgtggtct tccagtattc agatggaaaa ctacagactg tggctgaaaa ggaagtaaaa
     3601 ggagctgtgt actctatggt ggagtttaat gggaagctgc ttgccagcat caatagcacg
     3661 gtgcggctct atgagtggac cacagaaaag gagcttcgca ctgagtgtaa tcactacaac
     3721 aacatcatgg ccctgtacct gaagaccaag ggcgacttca tcctggtggg cgacctcatg
     3781 cgctcagtgc tgctgcttgc ctacaagcct atggaaggaa actttgaaga gattgcccga
     3841 gactttaatc ccaactggat gagtgcagtg gagatcttgg atgatgacaa ttttctggga
     3901 gctgaaaatg cctttaactt gtttgtgtgt cagaaggaca gtgctgccac cactgatgag
     3961 gagcggcagc acctacagga ggttggtctc ttccacctgg gcgagtttgt caatgtgttc
     4021 tgccatggct ccctagttat gcagaatctg ggcgaggctt ctacccctac acagggctcg
     4081 gtgctctttg gcacagtcaa cggcatgata gggctggtga cctcgctgtc agagagctgg
     4141 tataacctcc tgttggacat gcagaaccga ctcaataagg tcatcaaaag cgtgggcaag
     4201 attgagcact ccttctggag atcctttcac actgagcgaa agacagaacc agccacaggc
     4261 ttcatcgatg gtgatctgat tgaaagtttc ctagatatca gccgccctaa gatgcaggag
     4321 gttgtggcaa acttgcagta tgatgatggc agtggtatga agcgggaggc aactgcagat
     4381 gacctcatca aagtcgtgga ggagctaact cggatccatc tcgaggacta caaagaccat
     4441 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg
     4501 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4561 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4621 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4681 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4741 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4801 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4861 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4921 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4981 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5041 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5101 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5161 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5221 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5281 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5341 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5401 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5461 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5521 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5581 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5641 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5701 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5761 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5821 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5881 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5941 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6001 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6061 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6121 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6181 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6241 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6301 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6361 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6421 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6481 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6541 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6601 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6661 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6721 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6781 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6841 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6901 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6961 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7021 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7081 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7141 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7201 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7261 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7321 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7381 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7441 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7501 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7561 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7621 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7681 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     7741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7981 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8041 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8341 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8761 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8821 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8881 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
