LOCUS       Exported                7336 bp DNA     circular SYN 21-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7336)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2136..2534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2757..3527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(749..1204)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     terminator      complement(1332..1380)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     terminator      complement(1442..1490)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     promoter        complement(1697..1916)
                     /label=Hpa II promoter
                     /note="The HpaII promoter was a strong promoter in 
                     Gram-negative bacteria and  B. subtilis."
     CDS             complement(2136..2534)
                     /codon_start=1
                     /label=bleomycin binding protein
                     /translation="MLQSIPALPVGDIKKSIGFYCDKLGFTLVHHEDGFAVLMCNEVRI
                     HLWEASDEGWRSRSNDSPVCTGAESFIAGTASCRIEVEGIDELYQHIKPLGILHPNTSL
                     KDQWWDERDFAVIDPDNNLISFFQQIKS"
     CDS             complement(2757..3527)
                     /codon_start=1
                     /label=kanamycin nucleotidyltransferase
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG
                     QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
                     SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     CDS             complement(3687..4691)
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     promoter        5230..5332
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5333..5992
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTV*LDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      6080..6105
                     /label=trp terminator
     terminator      6108..6133
                     /label=trp terminator
     CDS             6307..7167
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      721 gccgcagccg aacgaccgag cgcagatcaa attgtaagcg ttaatatttt gttaaaattc
      781 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
      841 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
      901 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
      961 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     1021 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1081 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1141 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     1201 gcgtgatctg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     1261 aggttttcac cgtcatcacc gaaacgcgcg aggcagagct tggaattgcg aataataatt
     1321 ttttcacgtt gaaaatctcc aaaaaaaaag gctccaaaag gagcctttaa ttgtatcggt
     1381 ttatcagctt gctttcgagg tgaattagct tggaattgcg aataataatt ttttcacgtt
     1441 gaaaatctcc aaaaaaaaag gctccaaaag gagcctttaa ttgtatcggt ttatcagctt
     1501 gctttcgagg tgaatttcga cctctagaac gcgtgatctc agatctggta cgtaccaagc
     1561 tagcttgagc tcgactctag aggatccccc tgatttcact ttttgcattc tacaaactgc
     1621 ataactcata tgtaaatcgc tcctttttag gtggcacaaa tgtgaggcat tttcgctctt
     1681 tccggcaacc acttccaagt aaagtataac acactatact ttatattcat aaagtgtgtg
     1741 ctctgcgagg ctgtcggcag tgccgaccaa aaccataaaa cctttaagac ctttcttttt
     1801 tttacgagaa aaaagaaaca aaaaaacctg ccctctgcca cctcagcaaa ggggggtttt
     1861 gctctcgtgc tcgtttaaaa atcagcaagg gacaggtagt attttttgag aagatcactc
     1921 aaaaaatctc cacctttaaa cccttgccaa tttttatttt gtccgttttg tctagcttac
     1981 cgaaagccag actcagcaag aataaaattt ttattgtctt tcggttttct agtgtaacgg
     2041 acaaaaccac tcaaaataaa aaagatacaa gagaggtctc tcgtatcttt tattcagcaa
     2101 tcgcgcccga ttgctgaaca gattaataat agattttagc tttttatttg ttgaaaaaag
     2161 ctaatcaaat tgttgtcggg atcaattact gcaaagtctc gttcatccca ccactgatct
     2221 tttaatgatg tattggggtg caaaatgccc aaaggcttaa tatgttgata taattcatca
     2281 attccctcta cttcaatgcg gcaactagca gtaccagcaa taaacgactc cgcacctgta
     2341 caaaccggtg aatcattact acgagagcgc cagccttcat cacttgcctc ccatagatga
     2401 atccgaacct cattacacat tagaactgcg aatccatctt catggtgaac caaagtgaaa
     2461 cctagtttat cgcaataaaa acctatactc tttttaatat ccccgactgg caatgccggg
     2521 atagactgta acattctcac gcataaaatc ccctttcatt ttctaatgta aatctattac
     2581 cttattatta attcaattcg ctcataatta atcctttttc ttattacgca aaatggcccg
     2641 atttaagcac accctttatt ccgttaatgc gccatgacag ccatgataat tactaatact
     2701 aggagaagtt aataaatacg taaccaacat gattaacaat tattagaggt catcgttcaa
     2761 aatggtatgc gttttgacac atccactata tatccgtgtc gttctgtcca ctcctgaatc
     2821 ccattccaga aattctctag cgattccaga agtttctcag agtcggaaag ttgaccagac
     2881 attacgaact ggcacagatg gtcataacct gaaggaagat ctgattgctt aactgcttca
     2941 gttaagaccg aagcgctcgt cgtataacag atgcgatgat gcagaccaat caacatggca
     3001 cctgccattg ctacctgtac agtcaaggat ggtagaaatg ttgtcggtcc ttgcacacga
     3061 atattacgcc atttgcctgc atattcaaac agctcttcta cgataagggc acaaatcgca
     3121 tcgtggaacg tttgggcttc taccgattta gcagtttgat acactttctc taagtatcca
     3181 cctgaatcat aaatcggcaa aatagagaaa aattgaccat gtgtaagcgg ccaatctgat
     3241 tccacctgag atgcataatc tagtagaatc tcttcgctat caaaattcac ttccaccttc
     3301 cactcaccgg ttgtccattc atggctgaac tctgcttcct ctgttgacat gacacacatc
     3361 atctcaatat ccgaataggg cccatcagtc tgacgaccaa gagagccata aacaccaata
     3421 gccttaacat catccccata tttatccaat attcgttcct taatttcatg aacaatcttc
     3481 attctttctt ctctagtcat tattattggt ccattcacta ttctcatccc cttttcagat
     3541 aattttagat ttgcttttct aaataagaat atttggagag caccgttctt attcagctat
     3601 taataactcg tcttcctaag catccttcaa tccttttaat aacaattata gcatctaatc
     3661 ttcaacaaac tggcccgttt gttgaactac tctttaataa aataattttt ccgttcccaa
     3721 ttccacattg caataataga aaatccatct tcatcggctt tttcgtcatc atctgtatga
     3781 atcaaatcgc cttcttctgt gtcatcaagg tttaattttt tatgtatttc ttttaacaaa
     3841 ccaccatagg agattaacct tttacggtgt aaaccttcct ccaaatcaga caaacgtttc
     3901 aaattctttt cttcatcatc ggtcataaaa tccgtatcct ttacaggata ttttgcagtt
     3961 tcgtcaattg ccgattgtat atccgattta tatttatttt tcggtcgaat catttgaact
     4021 tttacatttg gatcatagtc taatttcatt gcctttttcc aaaattgaat ccattgtttt
     4081 tgattcacgt agttttctgt attcttaaaa taagttggtt ccacacatac caatacatgc
     4141 atgtgctgat tataagaatt atctttatta tttattgtca cttccgttgc acgcataaaa
     4201 ccaacaagat ttttattaat ttttttatat tgcatcattc ggcgaaatcc ttgagccata
     4261 tctgacaaac tcttatttaa ttcttcgcca tcataaacat ttttaactgt taatgtgaga
     4321 aacaaccaac gaactgttgg cttttgttta ataacttcag caacaacctt ttgtgactga
     4381 atgccatgtt tcattgctct cctccagttg cacattggac aaagcctgga tttacaaaac
     4441 cacactcgat acaactttct ttcgcctgtt tcacgatttt gtttatactc taatatttca
     4501 gcacaatctt ttactctttc agccttttta aattcaagaa tatgcagaag ttcaaagtaa
     4561 tcaacattag cgattttctt ttctctccat ggtctcactt ttccactttt tgtcttgtcc
     4621 actaaaaccc ttgatttttc atctgaataa atgctactat taggacacat aatattaaaa
     4681 gaaaccccca tctatttagt tatttgttta gtcacttata actttaacag atggggtttt
     4741 tctgtgcaac caattttaag ggttttcaat actttaaaac acatacatac caacacttca
     4801 acgcaccttt cagcaactaa aataaaaatg acgttatttc tatatgtatc aagataagaa
     4861 agaacaagtt caaaaccatc aaaaaaagac accttttcag gtgctttttt tattttataa
     4921 actcattccc tgatctcgac ttcgttcttt ttttacctct cggttatgag ttagttcaaa
     4981 ttcgttcttt ttaggttcta aatcgtgttt ttcttggaat tgtgctgttt tatcctttac
     5041 cttgtctcca agcttcttct agaggtaccg catgcgatat cgagctctcc cgggaattcc
     5101 ccgcgcgcga tgccctttcg tcttcgaata aatacctgtg acggaagatc acttcgcaga
     5161 ataaataaat cctggtgtcc ctgttgatac cgggaagccc tgggccaact tttggcgaaa
     5221 atgagacgtt gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact
     5281 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
     5341 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     5401 ggcatttcag tcagttgctc aatgtaccta taaccagacc gtttagctgg atattacggc
     5461 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     5521 tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt
     5581 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     5641 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     5701 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     5761 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggcgaatat
     5821 ggacaacttc ttcgcccccg ttttcactat gggcaaatat tatacgcaag gcgacaaggt
     5881 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     5941 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta
     6001 aggcagttat tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg ataataagcg
     6061 gatgaatggc agaaattcga gcccgcctaa tgagcgggct tttttttagc ccgcctaatg
     6121 agcgggcttt tttttcgaaa gcaaattcga cccatcgcgc gcggggagtc aactcagcaa
     6181 aagttcgatt tattcaacaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac
     6241 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataata ttgaaaaagg
     6301 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6361 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6421 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6481 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6541 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6601 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6661 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6721 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6781 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6841 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6901 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6961 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7021 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7081 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7141 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7201 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7261 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7321 aagatcaaag gatctt
//
