LOCUS       sequence-333625-        3667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LacI expression cassette (A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3667)
  AUTHORS   Greco FV, Pandi A, Erb TJ, Grierson CS, Gorochowski TE
  TITLE     Harnessing the central dogma for stringent multi-level control of 
            gene expression.
  JOURNAL   Nat Commun. 2021 Mar 19;12(1):1738. doi: 10.1038/s41467-021-21995-7.
  PUBMED    33741937
REFERENCE   2  (bases 1 to 3667)
  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: "Nat 
            Commun."; date: "2021-03-19"; volume: "12(1):1738. doi"; pages: " 
            10.1038/s41467-021-21995-7"
COMMENT     Sequence Label: pA-01
FEATURES             Location/Qualifiers
     source          1..3667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(68..1150)
                     /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="MKPVTLYDVAEYAGVSYMTVSRVVNQASHVSAKTREKVEAAMVEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIYSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1112..1131
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1424..1441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1534..2122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1614..1633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2293..3084,3085..3153))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2916..2935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3154..3258)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3559..3581
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3573..3590
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3574..3590
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3601..3620
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3601..3619
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcagtg aagtgaaaag aaaaaaggcc gcagagcggc ctttttagtt agatcggacc
       61 ccacatctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      121 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
      181 ccagtgagac tggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
      241 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
      301 ggatataaca tgagctatct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
      361 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
      421 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
      481 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
      541 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
      601 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtcct
      661 catgggagta aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
      721 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
      781 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
      841 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
      901 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
      961 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     1021 ccaccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     1081 ccacgcggga aacggtcata taagagacac cggcatactc tgcgacatcg tataacgtta
     1141 ctggtttcat attcacctcc atgaattgac tctcttccgg gcgctatcat gccataccgc
     1201 gaaaggtttt tcaccattcg atggcgcgcc gcggctggga atctgagaca tgagtcaaga
     1261 tatttgctcg gtaacgtatg ctggagtgag accgagctcg gagcacaaga ctggcctcat
     1321 gggccttccg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaac
     1381 atggtcatag ctgtttcctt gcgtattggg cgctctccgc ttcctcgctc actgactcgc
     1441 tgcgctcggt cgttcgggta aagcctgggg tgcctaatga gcaaaaggcc agcaaaaggc
     1501 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1561 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1621 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1681 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1741 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1801 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1861 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1921 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1981 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2041 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2101 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2161 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2221 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2281 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2401 accatctggc cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt
     2461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2581 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     2641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2881 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3121 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3181 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3241 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     3301 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3361 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtggccgc
     3421 tacagggcgc tcccattcgc cattcaggct gcgcaactgt tgggaagggc gtttcggtgc
     3481 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     3541 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgacg
     3601 taatacgact cactataggg cgaattggcg gaaggccgtc aaggccacgt gtcttgtccg
     3661 cggtacc
//
