LOCUS       Exported                5928 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 5928)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        183..201
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    202..226
                     /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             271..924
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             952..969
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             979..996
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1012..1056
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1087..1101
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             1192..1215
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      1292..1339
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1376..1831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1923..2738)
                     /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      2860..3448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3634..3773
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3875..4066)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4638..5720)
                     /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(5721..5798)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
       61 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
      121 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgatctcg atcccgcgaa
      181 attaatacga ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat
      241 tttgtttaac tttaagaagg agatatacat atgtccccta tactaggtta ttggaaaatt
      301 aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag
      361 catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg
      421 gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc
      481 atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca
      541 gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca
      601 tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg
      661 aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat
      721 cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg
      781 gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat
      841 aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt
      901 ggtggtggcg accatcctcc aaaatcggat ggttcaacta gtggttctgg tcatcaccat
      961 caccatcact ccgcgggtct ggtgccacgc ggtagtactg caattggtat gaaagaaacc
     1021 gctgctgcta aattcgaacg ccagcacatg gacagcccag atctgggtac cggtggtggc
     1081 tccggtgatg acgacgacaa gagtcccatg gatatcgggg atccgaattc tgtacaggcc
     1141 ttggcgcgcc tgcaggcgag ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac
     1201 caccaccacc accactaatt gattaatacc taggctgcta aacaaagccc gaaaggaagc
     1261 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1321 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     1381 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1441 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1501 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1561 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1621 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1681 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1741 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1801 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     1861 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     1921 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1981 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2041 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2101 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2161 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     2221 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     2281 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     2341 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     2401 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     2461 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     2521 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2581 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     2641 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2701 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     2761 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     2821 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2881 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2941 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3001 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3061 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3121 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3181 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3241 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3301 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3361 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3421 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3481 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3541 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3601 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3661 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     3721 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     3781 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3841 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3901 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     3961 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4021 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     4081 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     4141 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     4201 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4261 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4321 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4381 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4441 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     4501 tcaacgacag gagcacgatc atgctagtca tgccccgcgc ccaccggaag gagctgactg
     4561 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     4621 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4681 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     4741 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     4801 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     4861 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     4921 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     4981 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     5041 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     5101 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     5161 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     5221 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     5281 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     5341 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     5401 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     5461 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     5521 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     5581 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     5641 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     5701 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     5761 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     5821 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     5881 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggc
//
