LOCUS       Exported                7211 bp DNA     circular SYN 21-MAY-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7211)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7211)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      251..299
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      537..992
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1018..1122
                     /label=AmpR promoter
     CDS             1123..1980
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2151..2742
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3030..3051
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        complement(3512..3614)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             4153..5154
                     /codon_start=1
                     /label=repB
                     /note="RepB replication protein"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     CDS             5317..6087
                     /codon_start=1
                     /product="kanamycin resistance"
                     /label=KanR
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG
                     QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
                     SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     CDS             6310..6708
                     /codon_start=1
                     /label=ble
                     /note="bleomycin binding protein"
                     /translation="MLQSIPALPVGDIKKSIGFYCDKLGFTLVHHEDGFAVLMCNEVRI
                     HLWEASDEGWRSRSNDSPVCTGAESFIAGTASCRIEVEGIDELYQHIKPLGILHPNTSL
                     KDQWWDERDFAVIDPDNNLISFFQQIKS"
     promoter        6928..7147
                     /label=Hpa II promoter
                     /note="The HpaII promoter was a strong promoter in 
                     Gram-negative bacteria and  B. subtilis."
ORIGIN
        1 catatcctca tatggtcagc atccgccgca gcttcgaagc gtatgtcgat gacatgaata
       61 tcattactgt tctgattcct gctgaacaaa aggaaatcat gacaccgccg gaattcgtcg
      121 acaagcttgg taccagatct gatatccctg cagggatcct ctagagtcga gctcaagcta
      181 gcttggtacg taccagatct gagatcccgg gttctagagg tcgaaattca cctcgaaagc
      241 aagctgataa accgatacaa ttaaaggctc cttttggagc cttttttttt ggagattttc
      301 aacgtgaaaa aattattatt cgcaattcca agctaattca cctcgaaagc aagctgataa
      361 accgatacaa ttaaaggctc cttttggagc cttttttttt ggagattttc aacgtgaaaa
      421 aattattatt cgcaattcca agctctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
      481 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg cagatcacgc
      541 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      601 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      661 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      721 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      781 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      841 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      901 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      961 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     1021 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1081 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1141 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1201 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1261 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1321 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1381 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1441 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1501 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1561 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1621 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1681 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1741 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1801 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1861 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1921 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1981 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2041 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2101 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2161 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2221 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2281 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2341 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2401 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2461 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2521 gtactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2581 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2641 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2701 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2761 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2821 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2881 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2941 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3001 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     3061 caggctttac actttatgct tccggcatat tctcaataaa ccctttaggg aaataggcca
     3121 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     3181 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     3241 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg
     3301 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     3361 ttttctttac ggtctttaaa aaggccgtaa tatccagcta aacggtctgg ttataggtac
     3421 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     3481 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     3541 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     3601 ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa
     3661 cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcga
     3721 agacgaaagg gcatcgcgcg cgggaaattc ccgggagagc tcgatatcgc atgcggtacc
     3781 tctagaagaa gcttggagac aaggtaaagg ataaaacagc acaattccaa gaaaaacacg
     3841 atttagaacc taaaaagaac gaatttgaac taactcataa ccgagaggta aaaaaagaac
     3901 gaagtcgaga tcagggaatg agtttataaa ataaaaaaag cacctgaaaa ggtgtctttt
     3961 tttgatggtt ttgaacttgt tctttcttat cttgatacat atagaaataa cgtcattttt
     4021 attttagttg ctgaaaggtg cgttgaagtg ttggtatgta tgtgttttaa agtattgaaa
     4081 acccttaaaa ttggttgcac agaaaaaccc catctgttaa agttataagt gactaaacaa
     4141 ataactaaat agatgggggt ttcttttaat attatgtgtc ctaatagtag catttattca
     4201 gatgaaaaat caagggtttt agtggacaag acaaaaagtg gaaaagtgag accatggaga
     4261 gaaaagaaaa tcgctaatgt tgattacttt gaacttctgc atattcttga atttaaaaag
     4321 gctgaaagag taaaagattg tgctgaaata ttagagtata aacaaaatcg tgaaacaggc
     4381 gaaagaaagt tgtatcgagt gtggttttgt aaatccaggc tttgtccaat gtgcaactgg
     4441 aggagagcaa tgaaacatgg cattcagtca caaaaggttg ttgctgaagt tattaaacaa
     4501 aagccaacag ttcgttggtt gtttctcaca ttaacagtta aaaatgttta tgatggcgaa
     4561 gaattaaata agagtttgtc agatatggct caaggatttc gccgaatgat gcaatataaa
     4621 aaaattaata aaaatcttgt tggttttatg cgtgcaacgg aagtgacaat aaataataaa
     4681 gataattctt ataatcagca catgcatgta ttggtatgtg tggaaccaac ttattttaag
     4741 aatacagaaa actacgtgaa tcaaaaacaa tggattcaat tttggaaaaa ggcaatgaaa
     4801 ttagactatg atccaaatgt aaaagttcaa atgattcgac cgaaaaataa atataaatcg
     4861 gatatacaat cggcaattga cgaaactgca aaatatcctg taaaggatac ggattttatg
     4921 accgatgatg aagaaaagaa tttgaaacgt ttgtctgatt tggaggaagg tttacaccgt
     4981 aaaaggttaa tctcctatgg tggtttgtta aaagaaatac ataaaaaatt aaaccttgat
     5041 gacacagaag aaggcgattt gattcataca gatgatgacg aaaaagccga tgaagatgga
     5101 ttttctatta ttgcaatgtg gaattgggaa cggaaaaatt attttattaa agagtagttc
     5161 aacaaacggg ccagtttgtt gaagattaga tgctataatt gttattaaaa ggattgaagg
     5221 atgcttagga agacgagtta ttaatagctg aataagaacg gtgctctcca aatattctta
     5281 tttagaaaag caaatctaaa attatctgaa aagggaatga gaatagtgaa tggaccaata
     5341 ataatgacta gagaagaaag aatgaagatt gttcatgaaa ttaaggaacg aatattggat
     5401 aaatatgggg atgatgttaa ggctattggt gtttatggct ctcttggtcg tcagactgat
     5461 gggccctatt cggatattga gatgatgtgt gtcatgtcaa cagaggaagc agagttcagc
     5521 catgaatgga caaccggtga gtggaaggtg gaagtgaatt ttgatagcga agagattcta
     5581 ctagattatg catctcaggt ggaatcagat tggccgctta cacatggtca atttttctct
     5641 attttgccga tttatgattc aggtggatac ttagagaaag tgtatcaaac tgctaaatcg
     5701 gtagaagccc aaacgttcca cgatgcgatt tgtgccctta tcgtagaaga gctgtttgaa
     5761 tatgcaggca aatggcgtaa tattcgtgtg caaggaccga caacatttct accatccttg
     5821 actgtacagg tagcaatggc aggtgccatg ttgattggtc tgcatcatcg catctgttat
     5881 acgacgagcg cttcggtctt aactgaagca gttaagcaat cagatcttcc ttcaggttat
     5941 gaccatctgt gccagttcgt aatgtctggt caactttccg actctgagaa acttctggaa
     6001 tcgctagaga atttctggaa tgggattcag gagtggacag aacgacacgg atatatagtg
     6061 gatgtgtcaa aacgcatacc attttgaacg atgacctcta ataattgtta atcatgttgg
     6121 ttacgtattt attaacttct cctagtatta gtaattatca tggctgtcat ggcgcattaa
     6181 cggaataaag ggtgtgctta aatcgggcca ttttgcgtaa taagaaaaag gattaattat
     6241 gagcgaattg aattaataat aaggtaatag atttacatta gaaaatgaaa ggggatttta
     6301 tgcgtgagaa tgttacagtc tatcccggca ttgccagtcg gggatattaa aaagagtata
     6361 ggtttttatt gcgataaact aggtttcact ttggttcacc atgaagatgg attcgcagtt
     6421 ctaatgtgta atgaggttcg gattcatcta tgggaggcaa gtgatgaagg ctggcgctct
     6481 cgtagtaatg attcaccggt ttgtacaggt gcggagtcgt ttattgctgg tactgctagt
     6541 tgccgcattg aagtagaggg aattgatgaa ttatatcaac atattaagcc tttgggcatt
     6601 ttgcacccca atacatcatt aaaagatcag tggtgggatg aacgagactt tgcagtaatt
     6661 gatcccgaca acaatttgat tagctttttt caacaaataa aaagctaaaa tctattatta
     6721 atctgttcag caatcgggcg cgattgctga ataaaagata cgagagacct ctcttgtatc
     6781 ttttttattt tgagtggttt tgtccgttac actagaaaac cgaaagacaa taaaaatttt
     6841 attcttgctg agtctggctt tcggtaagct agacaaaacg gacaaaataa aaattggcaa
     6901 gggtttaaag gtggagattt tttgagtgat cttctcaaaa aatactacct gtcccttgct
     6961 gatttttaaa cgagcacgag agcaaaaccc ccctttgctg aggtggcaga gggcaggttt
     7021 ttttgtttct tttttctcgt aaaaaaaaga aaggtcttaa aggttttatg gttttggtcg
     7081 gcactgccga cagcctcgca gagcacacac tttatgaata taaagtatag tgtgttatac
     7141 tttacttgga agtggttgcc ggaaagagcg aaaatgcctc acatttgtgc cacctaaaaa
     7201 ggagcgattt a
//
