LOCUS       pEnCMV-HCN3(human)      8309 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018935
VERSION     V018935
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             665..2986
                     /label="HCN3(NM_020897)"
                     /note="HCN3(NM_020897)"
                     /gene="HCN3"
     CDS             2996..3061
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3085..3673
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3739..3787
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3989..4417
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4431..4760
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4827..5621
                     /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    5797..5918
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5967..5983)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5991..6007
                     /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(6015..6045)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6060..6081
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6369..6957)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7128..7988)
                     /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(7989..8093)
                     /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 ttaagcttgc
      661 caccatggag gcagagcagc ggccggcggc gggggccagc gaaggggcga cccctggact
      721 ggaggcggtg cctcccgttg ctcccccgcc tgcgaccgcg gcctcaggtc cgatccccaa
      781 atctgggcct gagcctaaga ggaggcacct tgggacgctg ctccagccta cggtcaacaa
      841 gttctccctt cgggtgttcg gcagccacaa agcagtggaa atcgagcagg agcgggtgaa
      901 gtcagcgggg gcctggatca tccaccccta cagcgacttc cggttttact gggacctgat
      961 catgctgctg ctgatggtgg ggaacctcat cgtcctgcct gtgggcatca ccttcttcaa
     1021 ggaggagaac tccccgcctt ggatcgtctt caacgtattg tctgatactt tcttcctact
     1081 ggatctggtg ctcaacttcc gaacgggcat cgtggtggag gagggtgctg agatcctgct
     1141 ggcaccgcgg gccatccgca cgcgctacct gcgcacctgg ttcctggttg acctcatctc
     1201 ttctatccct gtggattaca tcttcctagt ggtggagctg gagccacggt tggacgctga
     1261 ggtctacaaa acggcacggg ccctacgcat cgttcgcttc accaagatcc taagcctgct
     1321 gaggctgctc cgcctctccc gcctcatccg ctacatacac cagtgggagg agatctttca
     1381 catgacctat gacctggcca gtgctgtggt tcgcatcttc aacctcattg ggatgatgct
     1441 gctgctatgt cactgggatg gctgtctgca gttcctggtg cccatgctgc aggacttccc
     1501 tcccgactgc tgggtctcca tcaaccacat ggtgaaccac tcgtggggcc gccagtattc
     1561 ccatgccctg ttcaaggcca tgagccacat gctgtgcatt ggctatgggc agcaggcacc
     1621 tgtaggcatg cccgacgtct ggctcaccat gctcagcatg atcgtaggtg ccacatgcta
     1681 cgccatgttc atcggccatg ccacggcact catccagtcc ctggactctt cccggcgtca
     1741 gtaccaggag aagtacaagc aggtggagca gtacatgtcc ttccacaagc tgccagcaga
     1801 cacgcggcag cgcatccacg agtactatga gcaccggtac cagggcaaga tgttcgatga
     1861 ggaaagcatc ctgggcgagc tgagcgagcc gcttcgcgag gagatcatta acttcacctg
     1921 tcggggcctg gtggcccaca tgccgctgtt tgcccatgcc gaccccagct tcgtcactgc
     1981 agttctcacc aagctgcgct ttgaggtctt ccagccgggg gatctcgtgg tgcgtgaggg
     2041 ctccgtgggg aggaagatgt acttcatcca gcatgggctg ctcagtgtgc tggcccgcgg
     2101 cgcccgggac acacgcctca ccgatggatc ctactttggg gagatctgcc tgctaactag
     2161 gggccggcgc acagccagtg ttcgggctga cacctactgc cgcctttact cactcagcgt
     2221 ggaccatttc aatgctgtgc ttgaggagtt ccccatgatg cgccgggcct ttgagactgt
     2281 ggccatggat cggctgctcc gcatcggcaa gaagaattcc atactgcagc ggaagcgctc
     2341 cgagccaagt ccaggcagca gtggtggcat catggagcag cacttggtgc aacatgacag
     2401 agacatggct cggggtgttc ggggtcgggc cccgagcaca ggagctcagc ttagtggaaa
     2461 gccagtactg tgggagccac tggtacatgc gccccttcag gcagctgctg tgacctccaa
     2521 tgtggccatt gccctgactc atcagcgggg ccctctgccc ctctcccctg actctccagc
     2581 caccctcctt gctcgctctg cttggcgctc agcaggctct ccagcttccc cgctggtgcc
     2641 cgtccgagct ggcccatggg catccacctc ccgcctgccc gccccacctg cccgaaccct
     2701 gcacgccagc ctatcccggg cagggcgctc ccaggtctcc ctgctgggtc cccctccagg
     2761 aggaggtgga cggcggctag gacctcgggg ccgcccactc tcagcctccc aaccctctct
     2821 gcctcagcgg gcaacaggcg atggctctcc tgggcgtaag ggatcaggaa gtgagcggct
     2881 gcctccctca gggctcctgg ccaaacctcc aaggacagcc cagcccccca ggccaccagt
     2941 gcctgagcca gccacacccc ggggtctcca gctttctgcc aacatgtaac tcgaggacta
     3001 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     3061 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     3121 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3181 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3241 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3301 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3361 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3421 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     3481 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     3541 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3601 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3661 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     3721 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     3781 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3841 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3901 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     3961 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     4021 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4081 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4141 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4201 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4261 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4321 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     4381 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4441 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4501 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4561 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4621 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4681 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4741 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4801 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4861 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4921 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4981 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     5041 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5101 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5161 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5221 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5281 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5341 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5401 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5461 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5521 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5581 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     5641 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5701 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5761 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5821 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5881 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5941 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6001 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6061 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6121 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6181 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6241 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6301 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6361 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6421 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6481 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6541 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     6601 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6661 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6721 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6781 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6841 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6901 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6961 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7021 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7081 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7141 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7201 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7261 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7321 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7381 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7441 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7501 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7561 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7621 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7681 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7741 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7801 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7861 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7921 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7981 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8041 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8101 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     8161 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     8221 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     8281 ccgacaattg catgaagaat ctgcttagg
//
