LOCUS       V023541                 9023 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023541
VERSION     V023541
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             779..3703
                     /label="Syne3(NM_001042699)"
                     /note="Syne3(NM_001042699)"
                     /gene="Syne3"
     CDS             3710..3775
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3799..4387
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4453..4501
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4703..5131
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5145..5474
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5541..6335
                     /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    6511..6632
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6681..6697)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6705..6721
                     /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(6729..6759)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6774..6795
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7083..7671)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7842..8702)
                     /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(8703..8807)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 gacgcagcag ccccaggagg actttgagag gagcgtggag gatgcccagg cctggatgaa
      841 ggtgatacag gagcagcttc aggtcaatga caacacgaag gggccccgag cggccctgga
      901 ggcaaggctt cgagagacag agaaaatctg ccagctggag tctgaaggaa tggtgaaggt
      961 ggaactggtc ctgcgggcgg cggaggccct cttggcaact tgccaggagg gccagaaacc
     1021 tgagatcctg gcccggctga gggatatcaa gtctcagtgg gaggagacgg tcacctacat
     1081 gacccactgc cacagtcgca tcgagtgggt gtggctgcac tggagtgagt acctgctggc
     1141 ccaggatgag ttttaccgct ggttccagaa gatggtggtc gcactggagc cccccgtgga
     1201 gctgcagctg ggcttgaagg agaagcaatg gcagctgagc cacgcccagg tgctgctgca
     1261 caacgtggac aatcaggctg tgctcctgga caggctgttg gaggaggcgg gctccctgtt
     1321 cagcaggatc ggagacccca gcgtggatga agatgcccag aagaggatga aggctgagta
     1381 cgatgccgtg aaggccagag cccagcgcag ggtggacctc ctggcccagg tggcccagga
     1441 ccatgagcag taccgggagg acgtgaatga gttccagctg tggctgaagg cggtggtgga
     1501 gaaggtgcac agctgtctgg ggcggaactg caagctggcc acagaacttc gtctctctac
     1561 gctgcaggac atcgccaagg atttccctag gggtgaggag tctctgaaaa gattggagga
     1621 acaggctgtg ggtgtcattc aaaacacctc tcccttgggt gcagagaaga tctcagggga
     1681 gctggaggag atgcggggtg tcctggagaa gctgagagtc ctctggaaag aggaggaagg
     1741 gaggctgcgg ggcctgctcc agtccagggg ggactgtgag cagcagatcc aacagctgga
     1801 ggcagaactg ggagacttca agaaaagcct tcagaggctg gcccaggagg gcttggagcc
     1861 cacggtgaag acagccacag aggatgagct ggtggcccag tggaggctgt tctcggggac
     1921 tcgggctgca ctggcttcag aggaaccccg tgtagaccgg ttacaaactc aactgaagaa
     1981 acttgtcacc ttcccggacc tgcagtcact ctctgacagc gtggtagcca ccattcagga
     2041 ataccaaagt atgaagggga agaataccag gctccacaat gcgacccggg cagagctgtg
     2101 gcagcgtttc cagcggcccc taaatgacct gcagctgtgg aaggccctgg cccagaggct
     2161 cctggacatc acggccagcc tgcctgacct ggcctccatt cacacctttc taccccagat
     2221 tgaggcggcc ctcacggaaa gctctcgcct gaaagagcag ctggcgatgc tgcagctgaa
     2281 gaccgacctg ctgggcagca tctttggcca ggagagagca gccaccctcc tggagcaggt
     2341 gacaagttct gtgagggaca gagacctact gcataacagc cttcttcagc ggaagagcaa
     2401 acttcagagc ctgcttgttc agcacaagga ctttggggtg gcttttgatc ccctaaacag
     2461 gaagctccta gacctccagg ccaggatcca agcagagaaa gggcttccga gggaccttcc
     2521 tggaaagcag gtccagctcc taaggttgca ggggctgcag gaagaggggc tggatctggg
     2581 gacacagatc gaggctgtga ggcctcttgc ccatgggaac tctaagcacc agcagaaagt
     2641 agaccagatc tcctgtgacc agcaagccct gcagaggtcc ctggaggatc tcgtggacag
     2701 gtgtcagcag aacgtacggg aacattgtac cttcagtcac aggctgtcgg agctgcagct
     2761 atggatcacc atggccacac agacattaga gtcacaccaa ggggatgtgc gtctgtggga
     2821 tgctgagtcc caagaggctg gactcgagac gctgctgtct gaaatcccag agaaagaggt
     2881 ccaggtgtcc ctgctccaag cactgggcca gcttgtgatg aagaagtctt ccccagaagg
     2941 ggcaaccatg gtccaggagg agctgaggaa gctgatggag tcttggcagg ccctgcggct
     3001 gctagaggag aacatgctga gtctcatgag aaaccagcag ctgcagagga cagaggtgga
     3061 cacggggaag aagcaggtgt tcaccaacaa catcccaaag gccggctttc tcatcaaccc
     3121 tcaggacccc attcccagga gacagcatgg ggcaaaccca ctggaaggac acgacctccc
     3181 tgaagatcat ccccagctcc tgagggactt tgaacagtgg ctgcaggcag aaaactccaa
     3241 gctacgtaga atcatcacaa tgagagtggc cacagccaag gacttgagga ccagagaggt
     3301 gaagctgcag gagctggagg cccgaatccc agaaggccag cacctctttg agaacctgct
     3361 tcgtctcagg ccggcaaggg acccctccaa cgagctggaa gatctgcgct accggtggat
     3421 gctgtacaag tccaagctca aggactctgg ccacctgctg accgagagtt ctccggggga
     3481 gctgactgca ttccagaaga gtcggaggca gaagcggtgg agtccctgct ctctcctaca
     3541 gaaagcatgc cgtgtggcac tgccattgca gctgttgctc ctgctctttc tgctgctgct
     3601 gttcctgctg ccggccggcg aggaggagcg cagctgcgcc ctggccaaca acttcgcccg
     3661 ctcctttgcg ctcatgcttc ggtacaatgg ccccccgccc accctcgagg actacaaaga
     3721 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     3781 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3841 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3901 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3961 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     4021 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4081 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4141 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     4201 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     4261 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4321 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4381 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4441 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     4501 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4561 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4621 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     4681 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4741 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4801 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     4861 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4921 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4981 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5041 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     5101 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     5161 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5221 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5281 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5341 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5401 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5461 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     5521 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5581 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5641 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5701 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5761 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5821 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5881 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5941 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6001 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6061 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6121 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6181 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6241 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6301 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     6361 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     6421 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     6481 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     6541 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6601 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     6661 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6721 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6781 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6841 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6901 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6961 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7021 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7081 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7141 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7201 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7261 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7321 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7381 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7441 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7501 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     7561 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7621 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7681 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7741 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7801 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7861 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7921 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7981 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8041 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8101 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8161 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8221 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8281 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8341 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8401 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8461 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8521 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8581 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8641 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8701 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8761 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8821 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     8881 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     8941 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     9001 attgcatgaa gaatctgctt agg
//
