LOCUS       V038582                 6163 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038582
VERSION     V038582
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             925..1914
                     /label="EXTL2(NM_001439)"
                     /note="EXTL2(NM_001439)"
                     /gene="EXTL2"
     CDS             1927..1992
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2027..2251
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2297..2725
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2739..3068
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3080..3679
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    3839..3960
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4009..4025)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4033..4049
                     /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(4057..4087)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4102..4123
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4411..4996)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5167..6027)
                     /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(6028..6132)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagctttcta gagaattcgc caccatgagg tgttgccaca tctgcaaact tcctgggaga
      961 gtaatgggga ttcgagtgct tcgattatct ttggtggtca tcctcgtatt attactggta
     1021 gctggtgctt tgactgcctt acttcccagt gttaaagaag acaagatgct catgttgcgt
     1081 agggaaataa aatcccaggg caagtccacc atggactcct ttactctcat aatgcagacg
     1141 tacaacagaa cagatctctt attgaaactt ttaaatcatt atcaggctgt accaaatctg
     1201 cacaaagtga ttgtggtatg gaacaatatt ggagagaagg caccagatga attatggaat
     1261 tctctagggc cccaccctat ccctgtgatc ttcaaacaac agacagcaaa caggatgaga
     1321 aatcgactcc aggtctttcc tgaactggaa accaatgcag tgttgatggt agatgatgac
     1381 acactcatca gcaccccaga ccttgttttt gctttctcag tttggcagca atttcctgat
     1441 caaattgtag gatttgttcc tagaaagcac gtctctactt catcaggtat ctacagttat
     1501 ggaagttttg aaatgcaagc accagggtct ggaaatggtg accagtactc tatggtgctg
     1561 attggagcct cattcttcaa tagcaaatat cttgaattat ttcagaggca acctgcagct
     1621 gtccatgctt tgatagatga tactcaaaac tgtgatgata ttgccatgaa ttttatcatt
     1681 gccaagcata ttggcaagac ttcagggata tttgtgaagc ctgtaaacat ggacaatttg
     1741 gaaaaagaaa ccaacagtgg ctattctgga atgtggcatc gagctgagca cgctctgcag
     1801 aggtcttatt gtataaataa gcttgttaat atctatgata gcatgccctt aagatactcc
     1861 aacattatga tttcccagtt tggttttcca tatgccaact acaaaagaaa aatactcgag
     1921 accggtgact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     1981 gacgatgaca agtgaccggt aagtttaaac cgctgatcag cctcgactgt gccttctagt
     2041 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2101 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2161 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2221 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2281 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2341 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2401 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2461 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2521 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2581 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2641 tattcttttg atttataagg gattttggcc atttcggcct attggttaaa aaatgagctg
     2701 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2761 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2821 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2881 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2941 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3001 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3061 tttgcaaaaa gctcccggga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     3121 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     3181 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     3241 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     3301 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     3361 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     3421 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     3481 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     3541 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     3601 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     3661 ccgcaagccc ggtgcctgat tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3721 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3781 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     3841 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3901 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3961 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4021 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4081 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4141 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4201 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4261 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4321 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4381 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4441 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4501 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4561 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4621 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4681 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4741 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4801 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4861 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4921 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     4981 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5041 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5101 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5161 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5221 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5281 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5341 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5401 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5461 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5521 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5581 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5641 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5701 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5761 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5821 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5881 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5941 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6001 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6061 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6121 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
