LOCUS       Exported                5225 bp DNA     circular SYN 22-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pNCMO2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5225)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 22, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     Created by Takara Bio Inc. http://www.takarabio.com
            Cat. No. HB112
FEATURES             Location/Qualifiers
     source          1..5225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..103
                     /label=P2 promoter
     protein_bind    104..125
                     /label=Lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    258..347
                     /label=Secretion signal peptide
     misc_feature    346..409
                     /label=MCS
                     /note="multiple cloning site"
     CDS             926..1930
                     /codon_start=1
                     /label=RepB
                     /note="RepB replication protein from Enterococcus faecalis 
                     plasmid pAM-alpha-1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     CDS             2099..2860
                     /codon_start=1
                     /label=NmR
                     /note="Neomycin resistance gene, selection marker in 
                     Brevibacillus"
                     /translation="VNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     promoter        3083..3187
                     /label=AmpR promoter
     CDS             3188..4048
                     /codon_start=1
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVPLIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4219..4807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgatccgaca taatggacag gtgaataacg aaccacgaaa aaaactttaa atttttttcg
       61 aaggcgccgc aacttttgat tcgctcaggc gtttaatagg atgtaattgt gagcggataa
      121 caattattct gcatggcttt cctgcgaaag gaggtgacac gcgcttgcag gattcgggct
      181 ttaaaaagaa agatagatta acaacaaata ttccccaaga acaatttgtt tatactagag
      241 gaggagaaca caaggtcatg aaaaaaagaa gggtcgttaa cagtgtattg cttctgctac
      301 tgctagctag tgcactcgca cttactgttg ctcccatggc tttcgctgca ggatccgtcg
      361 actctagact cgaggaattc ggtaccccgg gttcgaaatc gataagcttc ggcattatag
      421 tgcggaggct ttttcgcatg caggtaggga acaattacat tgtctttgat tgtaaaaatg
      481 ctgttgacag gacactaaat ggtgtcgtat tctcaaagta acaccatttg gtgtccaatt
      541 gcaagtcatt tggtaacctt aattggatca gacaaggtaa aggataaaac agcacaattc
      601 caagaaaaac acgatttaga acctaaaaag aacgaatttg aactaactca taaccgagag
      661 gtaaaaaaag aacgaagtcg agatcaggga atgagtttat aaaataaaaa aagcacctga
      721 aaaggtgtct ttttttgatg gttttgaact tgttctttct tatcttgata catatagaaa
      781 taacgtcatt tttattttag ttgctgaaag gtgcgttgaa gtgttggtat gtatgtgttt
      841 taaagtattg aaaaccctta aaattggttg cacagaaaaa ccccatctgt taaagttata
      901 agtgactaaa caaataacta aatagatggg ggtttctttt aatattatgt gtcctaatag
      961 tagcatttat tcagatgaaa aatcaagggt tttagtggac aagacaaaaa gtggaaaagt
     1021 gagaccttgg agagaaaaga aaatcgctaa tgttgattac tttgaacttc tgcatattct
     1081 tgaatttaaa aaggctgaaa gagtaaaaga ttgtgctgaa atattagagt ataaacaaaa
     1141 tcgtgaaaca ggcgaaagaa agttgtatcg agtgtggttt tgtaaatcca ggctttgtcc
     1201 aatgtgcaac tggaggagag caatgaaaca tggcattcag tcacaaaagg ttgttgctga
     1261 agttattaaa caaaagccaa cagttcgttg gttgtttctc acattaacag ttaaaaatgt
     1321 ttatgatggc gaagaattaa ataagagttt gtcagatatg gctcaaggat ttcgccgaat
     1381 gatgcaatat aaaaaaatta ataaaaatct tgttggtttt atgcgtgcaa cggaagtgac
     1441 aataaataat aaagataatt cttataatca gcacatgcat gtattggtat gtgtggaacc
     1501 aacttatttt aagaatacag aaaactacgt gaatcaaaaa caatggattc aattttggaa
     1561 aaaggcaatg aaattagact atgatccaaa tgtaaaagtt caaatgattc gaccgaaaaa
     1621 taaatataaa tcggatatac aatcggcaat tgacgaaact gcaaaatatc ctgtaaagga
     1681 tacggatttt atgaccgatg atgaagaaaa gaatttgaaa cgtttgtctg atttggagga
     1741 aggtttacac cgtaaaaggt taatctccta tggtggtttg ttaaaagaaa tacataaaaa
     1801 attaaacctt gatgacacag aagaaggcga tttgattcat acagatgatg acgaaaaagc
     1861 cgatgaagat ggattttcta ttattgcaat gtggaattgg gaacggaaaa attattttat
     1921 taaagagtag ttcaacaaac gggccagttt gttgaagatt agatgctata attgttatta
     1981 aaaggattga aggatgctta ggaagacgag ttattaatag ctgaataaga acggtgctct
     2041 ccaaatattc ttatttagaa aagcaaatct aaaattatct gaaaagggaa tgagaatagt
     2101 gaatggacca ataataatga ctagagaaga aagaatgaag attgtccatg aaattaagga
     2161 acgaatattg gataaatatg gggatgatgt taaggctatt ggtgtttatg gctctcttgg
     2221 tcgtcagact gatgggccct attcggatat tgagatgatg tgtgtcatgt caacagagga
     2281 agcagagttc agccatgaat ggacaaccgg tgagtggaag gtggaagtga attttgatag
     2341 cgaagagatt ctactagatt atgcatctca ggtggaatca gattggccgc ttacacatgg
     2401 tcaatttttc tctattttgc cgatttatga ttcaggtgga tacttagaga aagtgtatca
     2461 aactgctaaa tcggtagaag cccaaacgtt ccacgatgcg atttgtgccc ttatcgtaga
     2521 agagctgttt gaatatgcag gcaaatggcg taatattcgt gtgcaaggac cgacaacatt
     2581 tctaccatcc ttgactgtac aggtagcaat ggcaggtgcc atgttgattg gtctgcatca
     2641 tcgcatctgt tatacgacga gcgcttcggt cttaactgaa gcagttaagc aatcagatct
     2701 tccttcaggt tatgaccatc tgtgccagtt cgtaatgtct ggtcaacttt ccgactctga
     2761 gaaacttctg gaatcgctag agaatttctg gaatgggatt caggagtgga cagaacgaca
     2821 cggatatata gtggatgtgt caaaacgcat accattttga acgatgacct ctaataattg
     2881 ttaatcatgt tggttacgta tttattaact tctcctagta ttagtaatta tcatggctgt
     2941 catggcgcat taacggaata aagggtgtgc ttaaatcggg cctgcatgac gaaagggcct
     3001 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3061 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3121 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3181 gaagagtatg agtattcaac atttccgtgt cccccttatt cccttttttg cggcattttg
     3241 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3301 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3361 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3421 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3481 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3541 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3601 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3661 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3721 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3781 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3841 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3901 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3961 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4021 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4081 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4141 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4201 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4261 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4321 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4381 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4441 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4501 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4561 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4621 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4681 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4741 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4801 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccatgcag
     4861 gctgaataaa agatacgaga gacctctctt gtatcttttt tattttgagt ggttttgtcc
     4921 gttacactag aaaaccgaaa gacaataaaa attttattct tgctgagtct ggctttcggt
     4981 aagctagaca aaacggacaa aataaaaatt ggcaagggtt taaaggtgga gattttttga
     5041 gtgatcttct caaaaaatac tacctgtccc ttgctgattt ttaaacgagc acgagagcaa
     5101 aacccccctt tgctgaggtg gcagagggca ggtttttttg tttctttttt ctcgtaaaaa
     5161 aaagaaaggt cttaaaggtt ttatggtttt ggtcggcact gccgacagcc tcgcagagca
     5221 cacac
//
