LOCUS       Exported                5356 bp DNA     circular SYN 10-SEP-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5356)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        265..283
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    284..308
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     CDS             353..418
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             485..502
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      569..616
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      653..1108
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1200..2015)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2137..2725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2911..3050
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3152..3343)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4152..5234)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5235..5312)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
       61 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
      121 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
      181 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
      241 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
      301 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac atatgaaata
      361 cctgctgccg accgctgctg ctggtctgct gctcctcgct gcccagccgg cgatggccat
      421 ggatatcgga attaattcgg atccgaattc gagctccgtc gacaagcttg cggccgcact
      481 cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga
      541 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
      601 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc
      661 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
      721 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
      781 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
      841 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
      901 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
      961 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     1021 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1081 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     1141 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct
     1201 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     1261 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     1321 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     1381 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     1441 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     1501 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     1561 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     1621 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     1681 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
     1741 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     1801 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     1861 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     1921 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     1981 ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     2041 taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2101 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2161 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2221 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2281 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2341 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2401 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2461 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2521 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2581 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2641 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2701 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2761 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2821 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2881 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     2941 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3001 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     3061 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3121 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3181 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     3241 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     3301 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     3361 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     3421 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     3481 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
     3541 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
     3601 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     3661 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     3721 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     3781 acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct
     3841 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     3901 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc
     3961 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag
     4021 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     4081 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     4141 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4201 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     4261 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     4321 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     4381 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
     4441 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     4501 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     4561 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     4621 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     4681 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     4741 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     4801 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     4861 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     4921 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     4981 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     5041 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     5101 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     5161 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     5221 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     5281 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
     5341 ctcctgcatt aggaag
//
