LOCUS       V030147                 6477 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030147
VERSION     V030147
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..1157
                     /label="Id2(NM_010496)"
                     /note="Id2(NM_010496)"
                     /gene="Id2"
     CDS             1164..1229
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1253..1841
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    1907..1955
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2157..2585
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2599..2928
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2995..3789
                     /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    3965..4086
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4135..4151)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4159..4175
                     /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(4183..4213)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4228..4249
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4537..5125)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5296..6156)
                     /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(6157..6261)
                     /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 agccttcagt ccggtgaggt
      781 ccgttaggaa aaacagcctg tcggaccaca gcttgggcat ctcccggagc aaaaccccgg
      841 tggacgaccc gatgagtctg ctctacaaca tgaacgactg ctactccaag ctcaaggaac
      901 tggtgcccag catcccccag aacaagaagg tgaccaagat ggaaatcctg cagcacgtca
      961 tcgattacat cttggacctg cagatcgccc tggactcgca tcccactatc gtcagcctgc
     1021 atcaccagag acctggacag aaccaggcgt ccaggacgcc gctgaccacc ctgaacacgg
     1081 acatcagcat cctgtccttg caggcatctg aattcccttc tgagcttatg tcgaatgata
     1141 gcaaagtact ctgtggcgga tccgactaca aagaccatga cggtgattat aaagatcatg
     1201 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     1261 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1321 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1381 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1441 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     1501 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     1561 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     1621 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     1681 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     1741 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     1801 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     1861 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     1921 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     1981 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2041 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2101 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     2161 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2221 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2281 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     2341 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2401 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2461 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2521 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2581 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2641 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2701 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2761 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2821 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2881 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2941 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3001 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3061 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3121 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3181 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3241 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3301 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3361 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3421 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3481 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3541 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3601 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3661 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3721 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3781 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     3841 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3901 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3961 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4081 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     4741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6301 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     6361 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6421 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
