LOCUS       V046008                 7917 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046008
VERSION     V046008
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..2597
                     /label="Zfp276(NM_020497)"
                     /note="Zfp276(NM_020497)"
                     /gene="Zfp276"
     CDS             2604..2669
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2693..3281
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3347..3395
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3597..4025
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4039..4368
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4435..5229
                     /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    5405..5526
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5575..5591)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5599..5615
                     /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(5623..5653)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5668..5689
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5977..6565)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6736..7596)
                     /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(7597..7701)
                     /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 ccaccatgaa gcgggaccgc ctcggccgct
      781 tcctgtctcc cggaatagct cggcagcgcg ggggttcggg cggcggctgt ggaagcggtc
      841 ggactcgggg tcgcccttct cgcagcggcg gaacgagcgc tgacggggcg gctgctcagc
      901 tcagctgggg ctcgatgaca cgctcctgcg gggacacagg cgacgacggt acagacgaag
      961 ctggagcagg ccggactctc gctatggggc attgtcgact ctgccatggg aagttttcct
     1021 cacggagcct ccgaagcatc tctgacagag tgcctgggga gacttcagaa agactgtccc
     1081 caggagagcg tgtcttcatt agggacttcc agcgtctgct gggtgtcgct gtgcaccagg
     1141 acccagccct gcctcagtct gtctgcaaga actgctacac ccaattctat cagtgccaca
     1201 gccttctcag gaccttctta cagagagtca acgtctcccc agctggccag aggaagcctt
     1261 gtacaaaggt tggtgttcag cctaccacag tggcagagga gggagcatgt gtggctgacc
     1321 tgatcgcctc cagccctcgg tgtctgcacg gcttggtggg ctgggtgcat gagcatgcag
     1381 tcagctgtgg gtctctgccc agcctccaga ggacactgtc ctctgagtac tgcggcatca
     1441 tccaggctgt gtggggctgc gaccagggcc acgacttcac catggacaca gcctccagct
     1501 gcagagctct ctttttggac agtgcattag cagtgaaatg ggcgtggggc aaggacctgt
     1561 caccacggct ggcccagaac tcagagtcca accccacggg agctgcctct cggctctgcc
     1621 aggccagaga aacccaagtt ggatctgaga ccaagacact ccccagtgtg gatgtggccc
     1681 tgctgcactc acatggagac tctgtgggac ctgggctagg cccctgtacg cagccacact
     1741 tggctcccag tgaggcccca gggcaactgg gtgagacgca ggttccctcg tcaacctcgg
     1801 atgatcgggt aaaagacgag ttcagtgacc tttctgaggg agacttcctg agcgaagatg
     1861 agagtgataa gaaacagaca cctcagtcct cggatgagtc ctttgagcct tacccagaaa
     1921 agaaggtctc tggtaagaag agcgaaggca gagaagccaa gaggcctgaa gagcccaaga
     1981 tcagaaagaa gcctgggccc aagcccggat ggaagaaaaa gcttcgctgt gagagggagg
     2041 agctgcccac catctacaag tgtccttacc agggctgcac agctgtatac cgtggcgctg
     2101 atggcatgaa gaagcacatt aaggagcacc atgaggaggt ccgggaaagg ccctgcccac
     2161 accctggctg caacaaggta ttcatgatcg accgatacct tcagcgacac gtgaagctca
     2221 tccacacaga ggtccgcaat tacatctgtg acgagtgcgg gcagaccttc aagcagcgga
     2281 agcaccttct cgtccaccag atgcgccact cgggagccaa gccactgcag tgtgaggtct
     2341 gtgggttcca gtgcaggcaa cgagcatccc tcaagtacca catgaccaaa cacaaagcag
     2401 agactgagct ggacttcgcc tgtgaccagt gcggccgacg ctttgaaaag gcccacaacc
     2461 tcaacgtgca tatgtccatg gtccatcctc tgacccaggc ccaggacaga gccctgcctc
     2521 tggaagcaga gccgccacct gggcctctga gcccctctgg gaccatggag ggtcaggctg
     2581 tgaagccgga gcctaccgga tccgactaca aagaccatga cggtgattat aaagatcatg
     2641 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     2701 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2761 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2821 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2881 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2941 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3001 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3061 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     3121 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3181 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3241 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     3301 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     3361 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3421 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3481 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3541 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     3601 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3661 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3721 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     3781 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3841 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3901 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3961 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4021 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4081 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4141 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4201 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4261 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4321 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4381 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4441 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4501 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4561 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4621 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4681 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4741 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4801 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4861 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4921 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4981 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5041 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5101 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5161 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5221 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     5281 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5341 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5401 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5461 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5521 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5581 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5641 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5701 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5761 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5821 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5881 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5941 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6001 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6061 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6121 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     6181 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6241 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6301 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6361 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6421 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6481 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6541 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6601 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6661 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6721 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6781 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6841 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6901 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6961 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7021 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7081 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7141 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7201 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7261 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7321 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7381 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7441 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7501 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7561 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7621 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7681 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7741 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     7801 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     7861 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
