LOCUS       V044754                 7336 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044754
VERSION     V044754
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..2826
                     /label="Pkd2l2(NM_001163004)"
                     /note="Pkd2l2(NM_001163004)"
                     /gene="Pkd2l2"
     CDS             2833..2898
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2936..3160
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3206..3634
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3648..3977
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4044..4838
                     /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    5012..5133
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5182..5198)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5206..5222
                     /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(5230..5260)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5275..5296
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5584..6169)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6340..7200)
                     /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(7201..7305)
                     /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 accatgtctg aggcgacttg gtggtaccga ggagggactt caaaacatga cctgcattac
     1021 agaagggaag cggaggttaa caccacactc gaggagttgt tactctattt tattttctta
     1081 ataaatctat gcatattgac ttttgggatg gtcaatccac atatgtacta tttaaacaaa
     1141 gttatgtcgt ctctgtttgt ggacacttct ctacctgatg atgaaagaag cagctttagg
     1201 tccattcgga gcataactga gttttggaag ttcatggaag gacccctcat cgacggcttg
     1261 tactgggact cgtggtatgg caacaaacag ctgtacagtg tgaagaacag cagccgcatc
     1321 tactacgaga acgttcttct cggcatcccc agagtgcggc aactgcgagt ccgaaacaac
     1381 acttgcaagg tctacccagc tttccagtcc ctggtcagcg actgctacag caagtacaca
     1441 gtggaaaacg aagacttctc tgattttggc ctcaaacgca atccagaatg gacgcacaca
     1501 ccttcttccc gcactgcccc atggcactgg gggtttgttg gcgtataccg agatggagga
     1561 tatatagtca cgttatcaaa atcaaaatct gaaaccaaag ccaaatttgt tgaccttcga
     1621 ctgaacaact ggattagcag aggcaccagg gctgttttta ttgatttctc cctgtacaat
     1681 gctaatgtca acctgttttg catcatcagg ctgctggcag agttccctgc gacgggtggg
     1741 ctcctcacct cctggcagtt ctactctgtg aagctcctca gatacgtctc ctactacgac
     1801 tacttcattg cctcctgtga agtcatattt tgtatttttc tctttgtctt cataatacaa
     1861 gaactgagga aagtgaacga gtttaagtct gcctatttca gaagtgtctg gaactggctg
     1921 gagatgctgc tcctgctgct ctgttttctc gccgtgtctt tctatgcata ctgtaacatg
     1981 cagagctttc tcttgcttgg acagctgctg aaaaacactg acagctatcc cgacttttac
     2041 ttccttgcat actggcacat ttactataac aacgtaattg ctatcactat cttctttgca
     2101 tggataaaga tattcaagtt cataagcttc aatgagacaa tgtcgcagct gtcatcaaca
     2161 ctctcccgct gcatgaagga catcgtgggg ttcgccatca tgttcttcat catcttctct
     2221 gcttatgccc agttgggatt tctggttttt gggtcacagg ttgatgattt ttcaactttt
     2281 caaaattcca tatttgcaca atttcgaatt gtcctcgggg actttaattt tgctggcatc
     2341 cagcaggcca actggatctt ggggcccatc tacttcatca cgttcatctt cttcgtgttc
     2401 tttgtgctcc tgaacatgtt cttggcaata attaatgaca cctattctga agttaaggct
     2461 gattattcaa taggcagaag accagatttt gaacttggta aaataattca aaagagttgc
     2521 tttaatgttc tcgagaaact cagactcaag aaagctcaag ctaaagaaga aaagaaaatg
     2581 caaaccactg acttggccca gagagccaga agagacggct ttgatgaaag tgagatccaa
     2641 gaggcagagc agatgaaaag atggaaggaa aggcttgaaa aaaagtatta ttctacagaa
     2701 attcaagacg attatcagcc tgtcactcag caagaattcc gagagctctt tttatacgcg
     2761 gtggagcttg agaaggaatt acactatgtc agtttaaaac tgaaccaact gatgagaaag
     2821 ctgcacctcg aggactacaa agaccatgac ggtgattata aagatcatga catcgactac
     2881 aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg
     2941 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3001 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3061 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3121 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3181 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3241 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3301 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3361 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3421 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3481 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3541 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3601 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3661 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3721 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3781 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3841 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3901 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3961 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4021 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4081 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4141 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4201 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4261 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4321 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4381 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4441 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4501 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4561 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4621 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4681 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4741 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4801 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4861 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4921 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4981 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5041 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5101 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5161 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5221 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5281 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5341 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5401 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5461 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5521 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5581 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5761 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5941 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6001 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6061 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6121 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6181 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6241 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6301 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6361 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6421 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6481 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6541 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6601 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6661 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6721 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6781 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6841 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6901 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6961 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7021 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7081 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7141 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7201 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7261 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7321 agtgccacct gacgtc
//
