LOCUS       V045364                 9399 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045364
VERSION     V045364
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..4079
                     /label="Gucy2e(NM_008192)"
                     /note="Gucy2e(NM_008192)"
                     /gene="Gucy2e"
     CDS             4086..4151
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4175..4763
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4829..4877
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5079..5507
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5521..5850
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5917..6711
                     /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    6887..7008
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7057..7073)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7081..7097
                     /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(7105..7135)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7150..7171
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7459..8047)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8218..9078)
                     /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(9079..9183)
                     /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 ctcgaattcg ccaccatgag cgcttggctc ctgccagccg
      781 gagggcttcc cggcgccggg ttctgtgtcc ctgcgcggca gtctccgtcc agtttctcgc
      841 gggtcctgcg ctggccaagg cctgggctac cgggactcct gctactgcta ctgctcccat
      901 ctccttctgc cctctctgct gtgttcaaag tgggggtgct gggcccctgg gcttgcgacc
      961 ccatctttgc acgggcccga ccagacctgg ctgcgcgtct ggccgccaac cgcctgaatc
     1021 gtgactttgc tttagacggc ggcccccggt tcgaggttgc gctgctccca gagccctgcc
     1081 tgactccggg ctcactaggg gctgtgtcct ctgcgctgtc tcgagtctct ggcctggtgg
     1141 gtccggtgaa ccccgcagcc tgtcggccag ccgaactgtt ggctcaagaa gctggagtag
     1201 cgctggtgcc ctggggctgc cctggcacgc gggcggcggg tactacagcc ccggcggtga
     1261 cccccgctgc agatgctctc tacgtcctcc ttagagcatt ccgctgggcg cgcgtggccc
     1321 tgatcactgc accccaagac ctgtgggtgg aggcgggacg cgctctgtcc acagcactca
     1381 gggcccgggg tttgccagtt gccctagtga cttccatgga gacttcagac cggtctggag
     1441 cccgggaggc cctcgggagg atccgagatg ggcctagagt tagagtagtg atcatggtga
     1501 tgcactcggt gctgctgggc ggcgaggagc agcgctacct actggaagct gcagaagaac
     1561 tggctctgac tgatggctcc ctggttttcc tgcccttcga cacgcttcac tacgctttgt
     1621 ctccaggccc ggaggctctg gctgcatttg tcaacagctc ccagctccgc agggctcacg
     1681 atgcggtgct cacactcacg cgccgctgtc ctcctggagg cagcgtgcaa gacagcctgc
     1741 gcagggctca agaacaccag gaactgcccc ttgacctcaa cctgaagcag gtctctccgc
     1801 tgtttggcac catctatgat gctgtcttcc tgttggctgg gggcgtgaag agagcaagaa
     1861 cagcggtggg tggtggctgg gtgtcaggtg catctgtagc ccgccaagta cgggaagcac
     1921 aagtctctgg cttttgtggg gtcctgggaa gaaccgagga gccctccttt gtgctgctgg
     1981 acacagatgc atccggagaa cagttgttcg caacacacct gctagatcct gtcttaggct
     2041 ccctgcgttc tgcagggacc cccatgcact tccctagagg tggacctgcc ccgggaccag
     2101 acccttcctg ctggttcgat ccagatgtga tctgcaacgg aggggtggag ccaggcctgg
     2161 tctttgttgg cttcctcctg gtgataggga tgggactgac tggagccttc ttggctcatt
     2221 acttgaggca caggctgcta cacatgcaga tggcttccgg ccccaacaag atcatcttga
     2281 cgttggaaga tgttactttc ctccacccac cgggaggcag ctctcgaaag gtggtccagg
     2341 gaagtagatc cagtctggct acccggagcg catcagacat tcgcagtgtc cccagccagc
     2401 cccaagagag caccaacgtt ggcctctatg agggggactg ggtttggctg aagaagttcc
     2461 caggggaaca tcatatggct atcaggccag caacaaagac agccttctcc aagcttcgag
     2521 agctccggca tgagaatgtg gctctctact tgggactctt cctggcgggt acagcagaca
     2581 gccctgccac ccctggggag ggcatcttgg ctgtggtctc agagcactgt gctcggggtt
     2641 ccctccatga cctcctggcc cagagagaaa taaagctgga ctggatgttc aagtcttccc
     2701 tcctgctgga cctcatcaag ggaatgagat atctgcacca tcgcggtgtg gcccacggga
     2761 ggctcaagtc acggaattgc gtggtggacg ggaggttcgt gctcaaggtg acagatcatg
     2821 gccatgggcg actgctggaa gcgcaaaggg tgttaccgga acctcccagt gcagaggatc
     2881 agctatggac agccccagag cttcttcggg acccctccct ggagcgccgg ggaactctag
     2941 ctggtgatgt ctttagtctg gccatcatca tgcaggaggt cgtgtgccgc agcacccctt
     3001 atgccatgct ggaactaacg cccgaggaag taatacagag ggtgcggagc cctcctccac
     3061 tgtgtcggcc cttggtgtcc atggaccagg cacccatgga gtgcatccag ctgatgacac
     3121 aatgctgggc agagcatcca gaacttcggc cttccatgga cctcaccttt gacctgttca
     3181 agagcatcaa caagggccgg aagaccaaca tcatcgactc catgcttcgg atgctggagc
     3241 agtactctag taacctggag gatctgatcc gagaacgcac agaggagtta gagcaggaga
     3301 agcagaagac agacaggctg ctcacacaga tgctgcctcc atctgtggct gaggccctga
     3361 agatggggac atctgtggag cctgagtact ttgaagaggt gacactctac ttcagtgaca
     3421 tcgtgggctt taccaccatt tcagccatga gcgagcctat tgaggtggta gacctgctta
     3481 atgacctcta tactctcttc gatgccatca tcggtgccca tgatgtctat aaggtggaaa
     3541 caattggaga tgcatatatg gtggcctccg ggctgccgca gaggaacggg cagcggcacg
     3601 ctgcagagat tgccaacatg tcactggaca tcctcagtgc agtcggctcc ttccgcatgc
     3661 gccatatgcc cgaggtaccg gtgcgcatcc gcattggttt gcactcaggc ccgtgcgtgg
     3721 cgggtgtggt gggcctcacc atgcctcggt actgcctgtt cggggacacg gtcaacactg
     3781 cctcgagaat ggagtccact ggactgcctt accgcatcca cgttaacatg agcactgttc
     3841 ggattcttcg cgctctggac caaggcttcc agatggagtg tcgaggccgc acggagctga
     3901 agggcaaggg tattgaggac acgtactggc ttgtgggcag acttggcttc aacaagccca
     3961 ttcccaaacc acctgatctg cagccagggg ccagcaacca tggtatcagc ctgcaggaga
     4021 ttcccccaga gagacgcaag aagctggaga aagccaggcc aggccagttt actgggaagg
     4081 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     4141 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     4201 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4261 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4321 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4381 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4441 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4501 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4561 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     4621 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4681 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4741 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     4801 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     4861 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4921 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4981 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5041 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     5101 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5161 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5221 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5281 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5341 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5401 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     5461 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     5521 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5581 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5641 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5701 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5761 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5821 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5881 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5941 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6001 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6061 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6121 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6181 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6241 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6301 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6361 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6421 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     6481 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6541 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6601 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6661 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6721 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6781 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6841 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6901 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6961 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7021 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7081 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7141 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7201 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7261 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7321 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7381 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7441 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7501 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7561 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7621 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     7681 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7741 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7801 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7861 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7921 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7981 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8041 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8101 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8161 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8221 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8281 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8341 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8401 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8461 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8521 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8581 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8641 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8701 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8761 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8821 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8881 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8941 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9001 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9061 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9121 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9181 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     9241 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     9301 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     9361 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
