LOCUS       V027953                 9222 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027953
VERSION     V027953
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..3902
                     /label="SSH1(NM_018984)"
                     /note="SSH1(NM_018984)"
                     /gene="SSH1"
     CDS             3909..3974
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3998..4586
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4652..4700
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4902..5330
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5344..5673
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5740..6534
                     /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    6710..6831
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6880..6896)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6904..6920
                     /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(6928..6958)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6973..6994
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7282..7870)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8041..8901)
                     /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(8902..9006)
                     /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 ccaccatggc cctggtgacc ctgcagcgct
      781 cgcccacgcc cagcgccgcc tcctcctcgg ccagcaacag cgagttggag gctggcagcg
      841 aagaagatcg aaaattaaac ctcagcttaa gtgagagctt tttcatggtg aaaggcgcag
      901 ccctcttctt acaacaggga agcagccctc aaggccagcg gagtcttcag cacccccaca
      961 agcatgcagg tgatctgcct caacatcttc aggtgatgat caaccttctg cgttgcgaag
     1021 acagaatcaa gctggcagtg cgcctggaga gcgcctgggc ggaccgggtc cggtacatgg
     1081 tggtggtgta cagcagcggg cgccaggaca ccgaggagaa tatcttgctg ggagtggact
     1141 tttccagtaa ggaaagtaaa agctgcacca ttgggatggt tctccgactg tggagcgaca
     1201 cgaaaatcca ccttgatgga gatggtgggt tcagcgtgag cacagcagga aggatgcaca
     1261 tatttaagcc tgtgtctgtc caggccatgt ggtctgccct gcaggtgctt cacaaggcct
     1321 gcgaagtggc ccggaggcac aactacttcc ccgggggtgt agctctcatc tgggctacct
     1381 actatgagag ctgcatcagc tccgagcaga gctgcatcaa cgagtggaac gccatgcagg
     1441 acctggagtc tacgcggccc gactcccccg cgctatttgt ggacaagccc actgaagggg
     1501 aaaggaccga gcgcctcatc aaagccaagc tccgaagcat catgatgagc caggatctag
     1561 aaaatgtgac ttccaaagag attcgtaatg aattagagaa acagatgaat tgtaacttga
     1621 aggaactcaa ggaatttata gacaatgaga tgctacttat cttgggacag atggacaagc
     1681 cctcccttat cttcgatcat ctttatctcg gctctgaatg gaatgcatcc aatctggagg
     1741 aactgcaggg ctcaggggtt gattacattt taaatgttac cagagaaatc gataattttt
     1801 ttcctggctt atttgcatat cataacatcc gagtctacga tgaagagacc acagacctcc
     1861 tcgcccactg gaatgaagcg tatcatttta taaacaaagc gaagaggaac cattccaagt
     1921 gcctggtgca ttgcaaaatg ggcgtgagtc gctcggcctc cacagtcata gcctatgcaa
     1981 tgaaggaatt cggctggcct ctggaaaaag catataacta tgtaaagcag aagcgcagca
     2041 tcacgcgccc caacgcgggc tttatgaggc agctgtctga gtatgaaggc atcttggatg
     2101 caagcaaaca gcggcacaac aagctgtggc gtcagcagac agacagcagc ctccagcagc
     2161 ctgtggatga ccctgcagga cctggcgact tcttgccaga gaccccagat ggcaccccgg
     2221 aaagccagct gcccttcttg gatgatgccg cccagcccgg cttagggccc cccctcccct
     2281 gctgtttccg gcgactctca gacccccttc tgccttcccc tgaggatgaa actggcagct
     2341 tggtccacct ggaggatccg gagagggagg ctctgttgga ggaagctgct ccacctgcag
     2401 aggtgcacag gccggccaga cagccccagc aaggttccgg actctgtgag aaggatgtga
     2461 agaagaaact agagtttggg agtcccaaag gtcggagcgg ctccttgctg caggtggagg
     2521 agacggaaag ggaggagggc ctgggagcag ggaggtgggg gcagcttcca acccagctcg
     2581 atcaaaacct gctcaactcg gagaacctaa acaacaacag caagaggagc tgtcccaacg
     2641 gcatggagga tgatgctata tttgggatcc ttaacaaagt gaagccttcc tataaatcct
     2701 gtgccgactg catgtaccct acagccagcg gggctcctga ggcctccagg gagcgatgtg
     2761 aggaccccaa tgctcccgcc atctgcaccc agccagcctt cctaccccac atcacgtcct
     2821 cccctgtggc ccacttggcc agcaggtccc gtgttccgga gaagccagcc tctggcccaa
     2881 ccgaacctcc cccgttccta ccaccagcag gctccaggag ggcagacacc agtggccctg
     2941 gggctggagc tgccctagag ccaccagcca gccttttgga accttccaga gagaccccaa
     3001 aagtcctgcc aaagtccctc cttttgaaga attctcactg tgataagaac cctcccagca
     3061 cagaagtggt aataaaggaa gaatcgtcac ccaagaaaga tatgaagcca gccaaggacc
     3121 tgaggcttct gttcagtaat gaatctgaga agccgacaac caacagctac ctgatgcagc
     3181 accaggagtc catcattcag ctgcagaagg caggcttggt ccgcaagcac accaaagagc
     3241 tagagcggct gaagagcgtg cctgcagacc cagcacctcc ctccagggat ggccctgcca
     3301 gcaggctgga ggccagcatc cccgaggaga gccaggatcc agccgcgctc cacgagctgg
     3361 gccccctggt tatgcccagc caggccggga gtgatgagaa gtcagaggcc gcccccgctt
     3421 cattggaagg aggctcactg aagagccccc ctcctttctt ctaccgcctg gaccacacca
     3481 gtagtttctc aaaagacttt ctgaagacca tctgctacac ccccacctcc tcttccatga
     3541 gctccaacct gacccggagc tccagcagcg atagcatcca cagtgtccgt gggaagcccg
     3601 ggctggtgaa gcagcggaca caggagattg agacccggct ccggctggcg ggcctcaccg
     3661 tctcttcccc actgaagcgc tcacactctc ttgccaagct ggggagtctc accttctcaa
     3721 cggaagacct gtccagtgag gctgacccgt ccaccgtcgc tgactcccag gacaccactt
     3781 tgagtgaatc ttccttcttg catgagcccc agggaacccc gagggaccca gctgcaacct
     3841 ccaaaccatc agggaaaccc gccccagaaa acttaaagag cccttcgtgg atgagcaaaa
     3901 gcggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     3961 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     4021 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4081 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4141 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4201 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4261 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4321 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4381 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     4441 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     4501 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4561 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     4621 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4681 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4741 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4801 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4861 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4921 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4981 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5041 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     5101 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5161 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5221 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     5281 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     5341 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5401 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5461 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5521 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5581 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     5641 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5701 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5761 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5821 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5881 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5941 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6001 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6061 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6121 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6181 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6241 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6301 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6361 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6421 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6481 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6541 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6601 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6661 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6721 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6781 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6841 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6901 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6961 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7021 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7081 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7141 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7201 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7441 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     7501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7681 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8101 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8161 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8221 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8281 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8341 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8401 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8461 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8521 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8581 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8641 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8701 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8761 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8821 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8881 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8941 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9001 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     9061 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     9121 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     9181 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
