LOCUS       Exported                4133 bp DNA     circular SYN 19-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4133)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4133)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        193..222
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    230..246
                     /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    270..326
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      529..615
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      707..734
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        754..844
                     /label=AmpR promoter
     CDS             845..1657
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1834..2422
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2608..2748)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        2934..3011
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3012..4091
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgg aattcgagct cggtacccgg ggatcctcta
      301 gagtcgacct gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc
      361 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
      421 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
      481 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
      541 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
      601 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
      661 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
      721 acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa
      781 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      841 gagtatgagc catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat
      901 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
      961 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1021 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     1081 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     1141 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     1201 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     1261 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     1321 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     1381 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     1441 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     1501 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     1561 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     1621 taaattgcag tttcatttga tgctcgatga gtttttctaa taactgtcag accaagttta
     1681 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1741 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1801 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1861 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1921 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1981 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2041 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2101 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2161 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2221 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2281 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2341 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2401 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2461 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2521 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2581 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     2641 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     2701 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     2761 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     2821 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     2881 gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcatttac gttgacacca
     2941 tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     3001 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     3061 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     3121 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     3181 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     3241 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     3301 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     3361 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     3421 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     3481 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     3541 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     3601 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     3661 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     3721 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     3781 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     3841 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     3901 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     3961 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     4021 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4081 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc gcgaattgat ctg
//
