LOCUS       sequence-202620-        3264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BSMT1 in Ecoli off a T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3264)
  AUTHORS   Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, 
            Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ
  TITLE     BioBits Explorer: A modular synthetic biology education kit.
  JOURNAL   Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. 
            eCollection 2018 Aug.
  PUBMED    30083608
REFERENCE   2  (bases 1 to 3264)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2018-08-1"; volume: "4(8):eaat5105. doi"; pages: " 
            10.1126/sciadv.aat5105. eCollection 2018 Aug"
COMMENT     Sequence Label: pJL1 BSMT1
FEATURES             Location/Qualifiers
     source          1..3264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     501..520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     754..771
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    894..915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(930..935,936..953,954..960)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    968..984
                     /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)."
     primer_bind     973..995
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     992..1008
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     992..1008
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1044..1063
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1044..1062
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1094..1116
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2249..2267)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2263..2310
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2328..3143)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3051..3070
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3135..3154
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ggatcctgca gttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
       61 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      121 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
      181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      361 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
      421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      661 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      721 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      781 gaagctcgca cgccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      841 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
      901 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      961 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgaa
     1021 ttcagatctc gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc
     1081 ctctagaaat aattttgttt aactttaaga aggagatata catatggaag ttgttgaagt
     1141 tcttcacatg aatggaggaa atggagacag tagctatgca aacaattctt tggttcagca
     1201 aaaggtgatt ctcatgacaa agccaataac tgagcaagcc atgattgatc tctacagcag
     1261 cctctttcca gaaaccttat gcattgcaga tttgggttgt tctttgggag ctaacacttt
     1321 cttggtggtc tcacagcttg ttaaaatagt agaaaaagaa cgaaaaaagc atggttttaa
     1381 gtctccagag ttttattttc acttcaatga tcttcctggc aatgatttta atacactttt
     1441 tcagtcactg ggggcatttc aagaagattt gagaaagcat ataggggaaa gctttggtcc
     1501 atgttttttc agtggagtgc ctggttcatt ttatactaga cttttccctt ccaaaagttt
     1561 acattttgtt tactcctcct acagtctcat gtggctatct caggtgccta atgggattga
     1621 aaataacaag ggaaacattt acatggcaag aacaagccct ctaagtgtta ttaaagcata
     1681 ctacaagcaa tatgaaatag atttttcaaa ttttctcaag taccgttcag aggaattgat
     1741 gaaaggtgga aagatggtgt taacactcct aggtagagaa agtgaggatc ctactagcaa
     1801 agaatgctgt tacatttggg agcttctagc catggccctc aataagttgg ttgaagaggg
     1861 attgataaaa gaagagaaag tagatgcatt caatattcct caatacacac catcaccagc
     1921 agaagtaaag tacatagttg agaaggaagg atcattcacc attaatcgct tggaaacatc
     1981 aagagttcat tggaatgctt ctaataatga gaagaatggt ggttacaatg tgtcaaggtg
     2041 catgagagct gtggctgagc ctttgcttgt cagccacttt gacaaggaat tgatggattt
     2101 agtgttccac aagtacgaag agattgtttc tgattgcatg tccaaagaga atactgagtt
     2161 tataaatgtc atcatctcct tgaccaaaat aaattaagtc gaccggctgc taacaaagcc
     2221 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     2281 gcctctaaac gggtcttgag gggttttttg ctgaaagcca attctgatta gaaaaactca
     2341 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     2401 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     2461 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     2521 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     2581 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     2641 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     2701 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     2761 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     2821 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     2881 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     2941 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     3001 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     3061 catttatacc catataaatc agcatccatg ttggaattta atcgcggctt cgagcaagac
     3121 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     3181 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     3241 acaacgtgag atcaaaggat cttc
//
