LOCUS       V028406                 7782 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028406
VERSION     V028406
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..2462
                     /label="Zfp697(NM_001411512)"
                     /note="Zfp697(NM_001411512)"
                     /gene="Zfp697"
     CDS             2469..2534
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2558..3146
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3212..3260
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3462..3890
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3904..4233
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4300..5094
                     /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    5270..5391
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5440..5456)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5464..5480
                     /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(5488..5518)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5533..5554
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5842..6430)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6601..7461)
                     /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(7462..7566)
                     /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 ccaccatgga acaggaagat aatcagggtg
      781 tgtgtgaata ccagacttct gaagacagag gcatggattc tgacttggag aactctgagg
      841 accgagaagg ggacccagaa gaaagaggaa tgggctctaa tccatgggac acagaagata
      901 gaggccacct ggagcaggaa gtggactcca acccacagga tgatgatcta agaggggact
      961 cacgagagag ggacatagcg tccactgtct gttcagaggg gcggctgagt gaggaagaaa
     1021 gggctatcct ccgtgaggaa gaggacgacc agcctggtgt ggctgacatg gctctgttcc
     1081 caggactgtc agaatctgac agcatatccc ggagtccgcg gggagaggaa gatgaagagg
     1141 aggaagacga agaagaggaa agcgctgggg agaaccgctt gatagaggag gaagatccat
     1201 tgcctactcc cgtgcttcct tggaggcgtc acctctccct gggcggtcga caccggggtg
     1261 acaagcctgc tcaccgccgc ttccaccggc tccatcaccc catggccatg gacctcgggg
     1321 aactagacag cctgatggcc agtatcatgg acgcgcccac catctgccca gactgcgggg
     1381 agagctttag cccgggcgcc gcattcctgc agcaccagcg catccaccgc ctggcagagg
     1441 ccgccgccgt ggccagcttg gagcccttcg gtttggccgg cgagtgcgga ggagtggtgg
     1501 ggatgatggg catgggtatg ggcgtgggca tgggcgtggc cggcggcttc ggggcggggc
     1561 ccacgctggc ccggccccct cgcgaaaagc ccttccgctg cggggagtgc ggcaaaggct
     1621 tcagccgcaa cacctacctg accaaccacc tgcggctgca cacgggcgag cggcccaacc
     1681 tgtgcgccga ctgcggcaag agcttcagct ggcgcgccga cctgctcaag caccggcgcc
     1741 tgcacacggg cgagaagccc tacccgtgcc cggagtgcgg cgaagccttc agcctcagct
     1801 cgcacttgct cagccaccgg cgcgcgcacg cggcggcggg cggcggcgcg ggctcggcgg
     1861 gctcggcggc ggcgctgcgg cccttcgcat gcggggagtg cggcaagggg ttcgtgcgcc
     1921 gctcgcatct ggccaatcac cagcgcatcc acaccggcga gaagccgcac ggctgcggcg
     1981 agtgcggcaa gcgcttcagc tggcgctcgg acctggtgaa gcaccagcgc gtgcacacgg
     2041 gcgagaagcc ctacatgtgc tccgagtgcg gcgagacctt cagcgtgagc tcgcacctct
     2101 tcacgcacaa gcgcacgcac tcgggcgagc ggccctacgt gtgccgggag tgtggcaagg
     2161 gcttcggccg caactcgcac ctcgtcaacc acctgcgcgt gcacaccggc gagaagccct
     2221 tcggctgcgg ccagtgcgag aagcgcttca gcgacttctc cacgctcacg cagcaccagc
     2281 gcacgcacac cggcgagaag ccctacacat gtctcgagtg cggcaagagc ttcatccaga
     2341 gctcccacct catccgccac cggcgcatcc acaccggcaa caagcctcac aaatgcgcgg
     2401 gctgtggcaa gggcttccgc tacaagacgc accttgcaca gcaccagaag ttgcacttgt
     2461 gtggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     2521 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     2581 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2641 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2701 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2761 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2821 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2881 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2941 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3001 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3061 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3121 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     3181 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     3241 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3301 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3361 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3421 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3481 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3541 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3601 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     3661 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3721 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3781 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     3841 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3901 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3961 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4081 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4141 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4201 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4261 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4321 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4381 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4441 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4501 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4561 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4621 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4681 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4741 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4801 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4861 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4921 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4981 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5041 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5101 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5161 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5221 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5281 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5341 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5401 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5461 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5521 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5581 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5641 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6001 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6241 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6661 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6901 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7201 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7561 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7621 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7681 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     7741 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
