LOCUS       T7_MBP_TEV_ISCro        7531 bp    DNA     circular SYN 06-JAN-2026
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7531)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 6, 2026 from SnapGene 8.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..327
                     /label=T7 promoter
                     /note="note: promoter for bacteriophage T7 RNA polymerase"
     protein_bind    328..352
                     /label=lac operator
                     /note="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)."
     RBS             364..386
                     /label=RBS
                     /note="note: efficient ribosome binding site from
                     bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             393..1493
                     /label=MBP
                     /note="codon_start: 1 gene: malE (mutated) note: This
                     version of the gene does not encode a signal sequence, so 
                     MBP will remain in the cytosol. product: maltose binding 
                     protein from E. coli transl_table: 1 translation: 
                     MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPD
                      
                     IIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIY
                      
                     NKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKY
                      
                     DIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNI
                      
                     DTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVN
                      
                     KDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASG
                      RQTVDEALKDAQT"
     CDS             1548..1568
                     /label=TEV site
                     /note="codon_start: 1 product: tobacco etch virus (TEV)
                     protease recognition and cleavage site transl_table: 1 
                     translation: ENLYFQS"
     primer_bind     1555..1593
                     /label=oJFC28
     primer_bind     complement(1555..1574)
                     /label=oJFC024
     primer_bind     complement(1555..1574)
                     /label=oJFC025
     CDS             1575..2552
                     /label=ISCro4 (E coli codon optimised)
                     /label=ISCRo4 E coli optimised (IDT)
                     /note="codon_start: 1 transl_table: 1 translation: 
                     MEQELHFIGIDVSKAKLDVDVLRPDGRHRSKKFANTPKGHDELLRWLSGHRVAPAHICM
                      
                     EATSTYMEDVAAHLSDAGYTVSVINPALGKAFAQSEGLRSKTDAVDARMLAEFCRQKRP
                      
                     PAWEAPHPVERALRALVLRHQSLTDMHTQELNRLETAREVQRPSIDAHLLWLHAELKRI
                      
                     EKQIKDLTDDDPDMKHRRKLLESIPGIGEKTSAVLLAYTGLKERFTHARQFAAFAGLTP
                      
                     RRYESGSSVNRASRMSKAGHASLRRALYMPAMVAVSKTEWGRAFRDRLAGNGKKGKVII
                      GAMMRKLAQVAYGVLKSGVPFDASRHNPVAA"
     primer_bind     1575..1593
                     /label=oJFC025
     CDS             2571..2588
                     /label=6xHis
                     /note="codon_start: 1 product: 6xHis affinity tag
                     transl_table: 1 translation: HHHHHH"
     terminator      2655..2702
                     /label=T7 terminator
                     /note="note: transcription terminator for bacteriophage T7
                     RNA polymerase"
     rep_origin      2739..3194
                     /label=f1 ori
                     /note="direction: RIGHT note: f1 bacteriophage origin of 
                     replication; arrow indicates direction of (+) strand 
                     synthesis"
     promoter        3220..3324
                     /label=AmpR promoter
                     /note="gene: bla"
     CDS             3325..3393
                     /label=AmpR
                     /note="codon_start: 1 gene: bla note: confers resistance to
                     ampicillin, carbenicillin, and related antibiotics product:
                     beta-lactamase transl_table: 1 translation: 
                     MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESF
                      
                     RPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTV
                      
                     RELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDER
                      
                     DTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGA
                      GERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     CDS             3394..4185
                     /label=AmpR
                     /note="codon_start: 1 gene: bla note: confers resistance to
                     ampicillin, carbenicillin, and related antibiotics product:
                     beta-lactamase transl_table: 1 translation: 
                     MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESF
                      
                     RPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTV
                      
                     RELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDER
                      
                     DTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGA
                      GERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     rep_origin      4356..4944
                     /label=ori
                     /note="direction: RIGHT note: high-copy-number 
                     ColE1/pMB1/pBR322/pUC origin of replication"
     misc_feature    5130..5269
                     /label=bom
                     /note="note: basis of mobility region from pBR322"
     CDS             complement(5371..5562)
                     /label=rop
                     /note="codon_start: 1 gene: rop product: Rop protein, which
                     maintains plasmids at low copy number transl_table: 1 
                     translation: 
                     VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDD
                      GENL*"
     primer_bind     5490..5510
                     /label=oJFC026
     primer_bind     complement(5490..5510)
                     /label=oJFC027
     CDS             complement(6371..7453)
                     /label=lacI
                     /note="codon_start: 1 gene: 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). product: lac
                     repressor transl_table: 1 translation: 
                     VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGK
                      
                     QSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQR
                      
                     VSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVAL
                      
                     GHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEG
                      
                     IVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRL
                      
                     LGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVS
                      RLESGQ*"
     promoter        complement(7454..7531)
                     /label=lacI promoter
                     /note="gene: lacI note: "
ORIGIN
        1 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
       61 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
      121 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
      181 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
      241 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
      301 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctgga
      361 tcctttgttt aactttaaga aggagatata ccatgaaaat cgaagaaggt aaactggtaa
      421 tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag aaattcgaga
      481 aagataccgg aattaaagtc accgttgagc atccggataa actggaagag aaattcccac
      541 aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac cgctttggtg
      601 gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc caggacaagc
      661 tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct tacccgatcg
      721 ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg ccaaaaacct
      781 gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc gcgctgatgt
      841 tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg ggttatgcgt
      901 tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac gctggcgcga
      961 aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat gcagacaccg
     1021 attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc atcaacggcc
     1081 cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg gtactgccga
     1141 ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt attaacgccg
     1201 ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg actgatgaag
     1261 gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag tcttacgagg
     1321 aagagttggc gaaagatcca cgtattgccg ccactatgga aaacgcccag aaaggtgaaa
     1381 tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact gcggtgatca
     1441 acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag actaattcga
     1501 gctcgaacaa caacaacaat aacaataaca acaacctcgg gatcgaggaa aacctgtact
     1561 tccaatccaa tgctatggag caagaactgc attttatcgg catagatgta tcgaaggcaa
     1621 aattagacgt tgatgtactt cgcccagacg gtagacatcg ctcgaagaaa ttcgccaata
     1681 ccccgaaggg ccacgacgaa ttgttgagat ggctgagtgg tcaccgcgta gcgccagctc
     1741 acatttgtat ggaagccacg tcgacttata tggaggatgt cgctgctcac cttagtgatg
     1801 cggggtacac tgtatcggtt atcaatcctg cacttggcaa ggccttcgct cagtcagaag
     1861 gattacgttc gaagacggac gctgttgatg cgcgtatgct ggcagagttc tgccggcaga
     1921 aacgccctcc ggcatgggag gcgcctcatc ccgtcgaacg tgcgctgaga gcgttggtac
     1981 tgcgccatca gtcgttgact gatatgcata cccaggagtt aaatcgcttg gagacggctc
     2041 gcgaagtgca gcgtcccagc atagatgctc atctgttgtg gttgcacgca gagttaaaac
     2101 gtattgaaaa acaaataaaa gacttgacag atgacgaccc cgacatgaag catcgccgta
     2161 agcttctgga atctatacct gggatcggcg aaaagacttc agctgttctt ttagcctata
     2221 ctggtctgaa ggagcgcttt acgcacgcgc gtcagttcgc cgcgttcgca gggttgacgc
     2281 ctcgccgcta tgaatcaggt tcaagcgtca atcgggctag ccgcatgagt aaagctgggc
     2341 acgcgtcgtt gcgcagagcc ttatacatgc ccgctatggt ggctgtgtcc aagactgaat
     2401 gggggcgggc ctttcgggat cgcttggcag ggaatgggaa gaaaggaaaa gtcattattg
     2461 gtgctatgat gcggaagctg gcgcaggtag cttatggagt cttgaagtcg ggagtcccct
     2521 tcgacgcctc gcgtcacaat cctgttgccg catccagcgg cagtctcgag caccaccacc
     2581 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     2641 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2701 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     2761 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2821 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2881 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2941 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3001 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3061 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3121 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3181 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3241 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3301 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3361 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3421 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3481 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3541 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3601 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3661 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3721 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3781 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3841 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     3901 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3961 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4021 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4081 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4141 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4201 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4261 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4321 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4381 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4441 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4501 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4561 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4621 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4681 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4741 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4801 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4861 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4921 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4981 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5041 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5101 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     5161 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5221 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     5281 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5341 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5401 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     5461 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     5521 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     5581 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     5641 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     5701 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     5761 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     5821 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
     5881 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     5941 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     6001 cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt
     6061 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     6121 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     6181 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     6241 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     6301 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     6361 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     6481 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     6541 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     6601 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     6661 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     6721 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     6781 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     6841 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     6901 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     6961 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     7021 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     7081 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     7141 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     7201 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     7261 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     7321 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     7381 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     7441 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     7501 cgcgaaaggt tttgcgccat tcgatggtgt c
//
