LOCUS       V038193                 6403 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038193
VERSION     V038193
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..1422
                     /label="REG1A(NM_002909)"
                     /note="REG1A(NM_002909)"
                     /gene="REG1A"
     CDS             1459..2154
                     /codon_start=1
                     /note=""
                     /product="promiscuous R40G mutant of a biotin protein
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /transl_table=1
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
                     /label="BioID2"
     CDS             2167..2232
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2267..2491
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2537..2965
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2979..3308
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3320..3919
                     /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    4079..4200
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4249..4265)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4273..4289
                     /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(4297..4327)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4342..4363
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4651..5236)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5407..6267)
                     /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(6268..6372)
                     /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 caccatggct cagaccagct catacttcat gctgatctcc
      961 tgcctgatgt ttctgtctca gagccaaggc caagaggccc agacagagtt gccccaggcc
     1021 cggatcagct gcccagaagg caccaatgcc tatcgctcct actgctacta ctttaatgaa
     1081 gaccgtgaga cctgggttga tgcagatctc tattgccaga acatgaattc gggcaacctg
     1141 gtgtctgtgc tcacccaggc cgagggtgcc tttgtggcct cactgattaa ggagagtggc
     1201 actgatgact tcaatgtctg gattggcctc catgacccca aaaagaaccg ccgctggcac
     1261 tggagcagtg ggtccctggt ctcctacaag tcctggggca ttggagcccc aagcagtgtt
     1321 aatcctggct actgtgtgag cctgacctca agcacaggat tccagaaatg gaaggatgtg
     1381 ccttgtgaag acaagttctc ctttgtctgc aagttcaaaa acggtggagg cggatcagga
     1441 ggcggtggat ctggatcctt caagaacctg atctggctga aggaggtgga cagcacccag
     1501 gagagactga aggagtggaa cgtgagctac ggcaccgccc tggtggccga cagacagacc
     1561 aagggcagag gcggcctggg cagaaagtgg ctgagccagg agggcggcct gtacttcagc
     1621 ttcctgctga accccaagga gttcgagaac ctgctgcagc tgcccctggt gctgggcctg
     1681 agcgtgagcg aggccctgga ggagatcacc gagatcccct tcagcctgaa gtggcccaac
     1741 gacgtgtact tccaggagaa gaaggtgagc ggcgtgctgt gcgagctgag caaggacaag
     1801 ctgatcgtgg gcatcggcat caacgtgaac cagagagaga tccccgagga gatcaaggac
     1861 agagccacca ccctgtacga gatcaccggc aaggactggg acagaaagga ggtgctgctg
     1921 aaggtgctga agagaatcag cgagaacctg aagaagttca aggagaagag cttcaaggag
     1981 ttcaagggca agatcgagag caagatgctg tacctgggcg aggaggtgaa gctgctgggc
     2041 gagggcaaga tcaccggcaa gctggtgggc ctgagcgaga agggcggcgc cctgatcctg
     2101 accgaggagg gcatcaagga gatcctgagc ggcgagttca gcctgagaag aagcctcgag
     2161 accggtgact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     2221 gacgatgaca agtgaccggt aagtttaaac cgctgatcag cctcgactgt gccttctagt
     2281 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2341 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2401 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2461 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2521 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2581 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2641 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2701 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2761 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2821 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2881 tattcttttg atttataagg gattttggcc atttcggcct attggttaaa aaatgagctg
     2941 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3001 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3061 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3121 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3181 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3241 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3301 tttgcaaaaa gctcccggga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     3361 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     3421 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     3481 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     3541 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     3601 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     3661 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     3721 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     3781 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     3841 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     3901 ccgcaagccc ggtgcctgat tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3961 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4021 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4141 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4201 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4261 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4321 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4381 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4441 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4501 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4561 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4621 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4681 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4741 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4801 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4861 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4921 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4981 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5041 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5101 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5161 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     5221 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5281 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5341 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5401 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5461 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5521 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5581 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5641 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5701 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5761 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5821 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5881 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5941 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6001 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6061 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6121 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6181 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6241 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6301 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6361 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
