LOCUS       V045989                 7113 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045989
VERSION     V045989
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..1793
                     /label="Enkd1(NM_198299)"
                     /note="Enkd1(NM_198299)"
                     /gene="Enkd1"
     CDS             1800..1865
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1889..2477
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2543..2591
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2793..3221
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3235..3564
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3631..4425
                     /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    4601..4722
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4771..4787)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4795..4811
                     /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(4819..4849)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4864..4885
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5173..5761)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5932..6792)
                     /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(6793..6897)
                     /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 ccaccatgtg cgagggcccg tcccgcatct
      781 ctggacccat ccccccagac cccactctct gcccggacta ctaccggcgg ccggcctcgg
      841 cccaaggacg cctggaagga aacgcgttga agctggacct gctgacttca ggtcgggacc
      901 tggactccag tcctcctcgt ggccctcgca tcaggcctga agcccgagag atcctagagc
      961 gtggccagcg aggcgtgggg gacgtgctgc tgcaactggg gtgcatctcc ctcggttcgg
     1021 gggtctctcc taagaggaag aatccaaagg accatgagaa ggaaaacttg aggcggatca
     1081 aagagattca gaggcgcttc caagaccagg aacgcagccg ggaacagggt cagcctaagc
     1141 cactgaaggc actgtggcgc tcacccaagt atgacaatgt ggagtctcga gtcaaggcca
     1201 ggatgaagga gcttggcccc acctctgtga cagaacctgc ccactttctg cgggcacact
     1261 cccgctgtgg ccctgggctc ccaccatccc gtgcctccag tcctcagctc gcccttccag
     1321 gtccccaagc taagggaccc ggcctaggcg tggatttcat tagtcgcaac gccctagctg
     1381 ccaagagagc cccccggcgc cactcccgct ccctgcaggt ccttgcacag gttcaggaac
     1441 agcagcgcca agcccaggag cgctacaacg ccacgcagaa aggccatgtg ccccattact
     1501 tgttggaacg cagggacctg tggcggaagg aagcggaagc tcgccagcgt agccagccag
     1561 acccgtccat gcccccgggc cacacactca tgcctgagaa tcagcggctg gagaccctga
     1621 acaatctgct acaaagccag agccagctgc tacgtgagct agtactatta cctgctgggg
     1681 cagactcgct gagagcccag ggtcatcgtg ctgaactgga ccggaagttg gtgcaaatag
     1741 aagaggctat caagatcttt tcccgaccca aagtctttgt gaagatggat accggatccg
     1801 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     1861 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     1921 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1981 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2041 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2101 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2161 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2221 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2281 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2341 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2401 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2461 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     2521 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     2581 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     2641 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     2701 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     2761 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2821 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2881 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2941 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3001 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3061 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3121 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     3181 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3241 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3301 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3361 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3421 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3481 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3541 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3601 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3661 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3721 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3781 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3841 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3901 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3961 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4021 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4081 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4141 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4201 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4261 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4321 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4381 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4441 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4501 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4561 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4621 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4681 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4741 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4801 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4861 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4921 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4981 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5041 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5101 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5161 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5221 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5281 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5341 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5401 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5461 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5521 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5581 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5641 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5701 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5761 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5821 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5881 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5941 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6001 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6061 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6121 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6181 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6241 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6301 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6361 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6421 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6481 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6541 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6601 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6661 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6721 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6781 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6841 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6901 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     6961 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     7021 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     7081 ttgaccgaca attgcatgaa gaatctgctt agg
//
