LOCUS       V046385                 7212 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046385
VERSION     V046385
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..1892
                     /label="Serpinb6c(NM_001382852)"
                     /note="Serpinb6c(NM_001382852)"
                     /gene="Serpinb6c"
     CDS             1899..1964
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1988..2576
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2642..2690
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2892..3320
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3334..3663
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3730..4524
                     /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    4700..4821
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4870..4886)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4894..4910
                     /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(4918..4948)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4963..4984
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5272..5860)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6031..6891)
                     /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(6892..6996)
                     /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 ccaccatgat ggatcctctg ctggaagcaa
      781 atgctacttt tgccttaaac cttttaaaaa tattgggtga agacagatcc aaaaatgtat
      841 ttttatcacc cataagcatc tcttcagcgc tggtcatggt cttattgggg gcaaaaggaa
      901 ccactgcgat ccagataact caggcactgt ctttgggtaa atgcagcagc agtgaagatg
      961 gagacgtcca ccagggcttc cagttacttc tcagcgaagt gaacaagact ggcacacagt
     1021 actcgctcaa agcagccaac aggctcttcg gggaaaagac ttttgatatc ctagcatctt
     1081 ttaaagattc ctgccacaag ttctatgaag cagagatgga agagctggac tttaagggtg
     1141 ctacagagca gtcccgacag cacatcaaca cctgggtagc caaaaagaca gaagataaaa
     1201 tcaaagagtt gctatctcca ggtacaatac attcaaacac tccactgatc ctcgtgaatg
     1261 ctgtctactt caaaggaaag tgggagaagc agtttaacaa agaggacacc agggagatgc
     1321 cattcaaagt cagcaagaat gaggagaaac ctgtgcaaat gatgtttcag aagtctacct
     1381 ttaagatgac ctatgtggaa gagatatcca ccaagatttt gttgcttccc tatgttggca
     1441 atgagctgaa catgatcatc atgcttccag atgaacacgt tgaactgagt acggtggaaa
     1501 aggaaataac tcatgagaaa tttatagagt ggacgaggct ggacaggatg aaaggagaaa
     1561 aggtggaggt tttcctccca tggtttaagc tggaggagaa ctatgacatg aaggatgtcc
     1621 tctgcaagct gggcatgact gatgcctttg aggagggcag agcagacttt tctggaatat
     1681 cttccaagca aggcttgttt ctgtccaatg tcatacataa atctgttgtg gaggtgaatg
     1741 aggaaggctc agaggctaca gctgccacta caattgttct gaaggggtca tcacggtcca
     1801 ccccttgttt ctgtgttaac cgccccttca ttttcttcat tcagcacatt aagaccaatg
     1861 agattctgtt ccttggccgg ctctcctctc ccggatccga ctacaaagac catgacggtg
     1921 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     1981 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2041 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2101 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2161 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2221 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2281 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2341 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     2401 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2461 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2521 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     2581 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     2641 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     2701 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     2761 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     2821 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     2881 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2941 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3001 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3061 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3121 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3181 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3241 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     3301 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3361 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3421 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3481 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3541 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3601 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3661 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3721 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3781 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3841 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3901 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3961 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4021 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4081 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4141 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4201 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4261 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4321 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4381 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4441 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4501 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     4561 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4621 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4681 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4741 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4801 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4861 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4921 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4981 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5041 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5101 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5161 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5221 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5281 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5341 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5401 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5461 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5521 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5581 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5641 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5701 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5761 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5821 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5881 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5941 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6001 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6061 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6121 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6181 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6241 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6301 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6361 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6421 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6481 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6541 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6601 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6661 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6721 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6781 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6841 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6901 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6961 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7021 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     7081 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7141 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     7201 aatctgctta gg
//
