LOCUS       V040748                 8042 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040748
VERSION     V040748
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"
     CDS             683..2722
                     /label="Zfp263(NM_148924)"
                     /note="Zfp263(NM_148924)"
                     /gene="Zfp263"
     CDS             2729..2794
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2818..3406
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3472..3520
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3722..4150
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4164..4493
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4560..5354
                     /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    5530..5651
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5700..5716)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5724..5740
                     /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(5748..5778)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5793..5814
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6102..6690)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6861..7721)
                     /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(7722..7826)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgactat ggctgcgggc ccaagctcgc aggagccaga
      721 agggctcctc atagtgaaac tggaggagga ctgtgcctgg agccacgagg tacccccgcc
      781 tgagccagag cccagcccgg aggcctccca tctgcgattt cgacggttcc gctttcagga
      841 cgccccgggt ccccgggagg cccttagccg gcttcaggag ctttgccgcg ggtggctgcg
      901 gccggagatg cgcaccaagg aacagatcct ggagctgttg gtgctggagc agttcttgac
      961 tatccttccc caggagattc agagcagggt gcaggagctg cgcccagaga gcggcgaaga
     1021 agcagtcact cttgtggagc gtatgcagaa agaacttggg aaactgaggc aacaggtcac
     1081 aaaccaaggg cggggagcag aagtgctttt ggaggagcct ttgccactgg aaacagcagg
     1141 agagtcaccg agcttcaagc tggagccaat ggagactgag cgaagccctg gccccaggct
     1201 gcaggagctg ctagacccca gcccccaaag ggactcccag gctgtaaagg agagggcatt
     1261 atccgctccc tggctctctc tcttccctcc tgaagggaat gtggaagata aggatatgac
     1321 tgggacccag ttgcccgaga gcctggagga catggcaatg tatatctctc aggagtggga
     1381 ccatcaggat cctagtaaga gagctctatc aaggtatatg gtacaagata gctatgagaa
     1441 ctcggggact ctagagtcca gcattcccag tcaggaggtc tcaagcaccc acgtggagca
     1501 aggagagaag ctgtgggatt caagtgtcca gacctgcaag gaaggcatga accccagaaa
     1561 cccagttcca ggagtggaaa agtttgagaa ccaagaaaga aatgttgagt ctgtttctcc
     1621 tgaaagtacc catcctccag tactattgcc tggccaggct agaagggagg tgccctggag
     1681 tcctgagcag ggaagacttg atgacagaga aggacattgg gaatgtcccc cagaggacaa
     1741 aatagaggag tctttagtgg gaaccccaag ttgtaaagga ctggtgcaag caaaagaaca
     1801 gcccaagaaa ctccatttgt gtgccttgtg tggcaagaac ttttctaata actcaaacct
     1861 aattaggcac cagagaatac atgcagcaga aaagctgtgt atggatgtag agtgtggtga
     1921 agtctttggt ggacacccac attttctgtc actgcacaga acacacatag gagaggaagc
     1981 tcataagtgc cttgaatgtg gaaaatgctt cagtcaaaat acccacctca cccgccatca
     2041 gcgcacccac acaggggaaa agcccttcca gtgcaatgcc tgtgggaaaa gcttctcttg
     2101 caactccaac ctcaacagac accaaaggac acacactggg gagaagccct acaagtgccc
     2161 agagtgtggg gagatctttg ctcacagctc caacctgctt cgtcaccaga ggatccacac
     2221 tggagagagg ccctatcgct gtagtgaatg tggcaaaagt ttctctcgga gttcacatct
     2281 tgttattcat gaaagaactc atgagaaaga gagactcgac cctttccctg agtgtggtca
     2341 aggcatgaat gacagtgccc cctttcttac aaaccacaga gtggagaaga aactctttga
     2401 atgttccact tgtgggaaaa gtttccggca gggcatgcac cttaccagac atcagagaac
     2461 acacacagga gaaaaaccgt ataagtgtat cctttgtggg gaaaacttct ctcatagatc
     2521 taacttaatc aggcaccaga ggattcatac aggagaaaaa ccctatacct gtcacgagtg
     2581 tggagacagt ttctctcaca gttccaaccg gattcgtcac ctgagaaccc atacaggaga
     2641 aagaccttat aaatgttctg aatgtggaga aagcttctct cggagctcac gtcttacaag
     2701 tcatcaaagg actcacactg gtctcgagga ctacaaagac catgacggtg attataaaga
     2761 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat
     2821 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2881 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2941 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3001 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3061 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3121 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3181 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3241 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3301 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3361 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     3421 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3481 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3541 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3601 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3661 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     3721 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3781 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3841 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     3901 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3961 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4021 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4081 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4141 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4201 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4261 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4321 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4381 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4441 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4501 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4561 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4621 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4681 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4741 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4801 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4861 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4921 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4981 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5041 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5101 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5161 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5221 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5281 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5341 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     5401 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5461 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5521 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5581 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5641 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5701 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5761 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5821 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5881 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5941 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6001 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6061 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6121 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6181 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6241 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6301 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6361 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6421 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6481 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6541 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6601 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6661 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6721 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6781 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6841 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6901 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6961 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7021 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7081 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7141 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7201 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7261 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7321 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7381 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7441 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7501 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7561 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7621 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7681 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7741 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7801 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     7861 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     7921 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     7981 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     8041 gg
//
