LOCUS       Exported                5367 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 5367)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        255..273
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    274..298
                     /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             346..390
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             406..423
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             482..499
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      566..613
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      650..1105
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1197..2012)
                     /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      2134..2722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2908..3047
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3149..3340)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4149..5231)
                     /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(5232..5309)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
       61 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
      121 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
      181 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatcgatct
      241 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
      301 tctagaaata attttgttta actttaagaa ggagatatac atatgaaaga aaccgctgct
      361 gctaaattcg aacgccagca catggacagc ccagatctgg gtaccctggt gccacgcggt
      421 tccatggctg atatcggatc cgaattcgag ctccgtcgac aagcttgcgg ccgcactcga
      481 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
      541 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
      601 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
      661 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
      721 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
      781 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
      841 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
      901 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
      961 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1021 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     1081 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1141 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag
     1201 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     1261 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     1321 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     1381 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     1441 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     1501 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     1561 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     1621 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     1681 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     1741 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     1801 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     1861 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     1921 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
     1981 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     2041 gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2101 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2161 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2221 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2281 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2341 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2401 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2461 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2521 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2581 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2641 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2701 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2761 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2821 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2881 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     2941 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3001 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     3061 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3121 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3181 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     3241 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     3301 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     3361 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     3421 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     3481 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     3541 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     3601 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     3661 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     3721 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     3781 acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct
     3841 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     3901 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     3961 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     4021 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     4081 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     4141 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4201 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     4261 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     4321 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     4381 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca
     4441 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     4501 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     4561 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     4621 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     4681 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     4741 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     4801 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     4861 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     4921 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     4981 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     5041 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     5101 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     5161 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     5221 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     5281 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     5341 ctgcattagg aagcagccca gtagtag
//
