LOCUS       sequence-324799-        2846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of LacI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2846)
  AUTHORS   Jung JK, Rasor BJ, Rybnicky GA, Silverman AD, Standeven J, Kuhn R, 
            Granito T, Ekas HM, Wang BM, Karim AS, Lucks JB, Jewett MC
  TITLE     At-Home, Cell-Free Synthetic Biology Education Modules for 
            Transcriptional Regulation and Environmental Water Quality 
            Monitoring.
  JOURNAL   ACS Synth Biol. 2023 Oct 20;12(10):2909-2921. doi: 
            10.1021/acssynbio.3c00223. Epub 2023 Sep 12.
  PUBMED    37699423
REFERENCE   2  (bases 1 to 2846)
  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"; doi: 
            "10.1021/acssynbio.3c00223"; journalName: "ACS Synth Biol"; date: 
            "2023-10-20- 20"; volume: "12"; issue: "10"; pages: "2909-2921"
COMMENT     Sequence Label: pJBL7084
FEATURES             Location/Qualifiers
     source          1..2846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     194..213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     321..340
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        321..339
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             371..393
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             401..1483
                     /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"
     primer_bind     complement(420..439)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1535..1553)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1549..1596
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1614..2429)
                     /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     2337..2356
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2421..2440
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2551..2846,1..293)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
       61 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      121 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      181 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      241 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      301 gcaacgcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
      361 tagaaataat tttgtttaac tttaagaagg agatatacat atgaaaccag taacgttata
      421 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
      481 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
      541 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
      601 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
      661 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
      721 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
      781 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
      841 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
      901 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
      961 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     1021 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     1081 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     1141 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     1201 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     1261 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     1321 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     1381 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1441 gcagctggca cgacaggttt cccgactgga aagcgggcag taagtcgacc ggctgctaac
     1501 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     1561 cttggggcct ctaaacgggt cttgaggggt tttttgctga aagccaattc tgattagaaa
     1621 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1681 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     1741 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     1801 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1861 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     1921 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     1981 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2041 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2101 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     2161 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2221 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2281 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2341 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcttcgag
     2401 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     2461 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     2521 tgagacacaa cgtgagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2581 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2641 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2701 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2761 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2821 gttggactca agacgatagt taccgg
//
