LOCUS       sequence-178390-        4682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:CelR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4682)
  TITLE     Swint-Kruse lab plasmids
REFERENCE   2  (bases 1 to 4682)
  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"
COMMENT     Sequence Label: LEhE-pHG165
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..82
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(102..121)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1319..1335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1319..1335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1332..1354)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1343..1359
                     /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)."
     promoter        complement(join(1367..1373,1374..1391,1392..1397))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1412..1433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2033..2224
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2218..2240)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2326..2466
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2381..2400
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2481..2498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2652..3240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2732..2751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3411..4202,4203..4271))
                     /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     4034..4053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4272..4376)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4444..4462
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4615..4637
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4629..4646
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4630..4646
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
       61 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
      121 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
      181 aacgcgggaa aaagtggaag cggcgatggc ggagctcggc tacgtgccga accgcgcagc
      241 ccgcaccctg gtcacccgac gtaccgacac cgtagccctg gtggtgtcgg aaaacaacca
      301 gaagctcttc gccgaaccct tctatgccgg gatcgtgctc ggcgtggggg ttgctctgtc
      361 cgaacgggga ttccagttcg tcctggccac gggccgctcc gggatagagc atgagcggct
      421 gggcggctac ctggccggac agcacgtcga cggggtcctc ctgctgtcgc tccaccgcga
      481 cgacccgctg ccgcagatgc tggacgaggc cggggtgccg tacgtctacg gcggccgtcc
      541 gctcggcgtc cccgaagaac aggtgtccta tgtcgatatc gacaacatcg gcgggggacg
      601 ccaggccacc cagcggctga tcgagaccgg gcaccggcgg atcgctacga tcgcgggccc
      661 gcaggacatg gtcgctggtg tggaacgcct ccaggggtat cgcgaagcac tgctcgccgc
      721 ggggatggag tacgacgaga cgctggtgag ctacggtgac ttcacctacg acagcggggt
      781 ggccgcgatg cgggagctgc tggatcgggc ccccgacgtg gacgccgtgt tcgcggcctc
      841 cgacttgatg gggctggccg cgctgcgggt gctgcgtgct tcgggacgcc gcgtgcccga
      901 ggatgtggcg gtggtcggct acgacgactc gaccgtagcc gagcacgccg aaccgccgat
      961 gaccagcgtc aaccagccca ccgagctgat gggccgggag atggcccggc tgctcgtgga
     1021 ccggatcacc ggggagacca ccgaaccggt gcggctggtg ctggagaccc atttgatggt
     1081 gcgggaatcc gggtgataac ctgaggccga tactgtcgtc gtcccctcaa actggcagat
     1141 gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattaggt caatccgccg
     1201 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
     1261 tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaattcg taatcatggt
     1321 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1381 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1441 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1501 gccaacgcgc ggggagaggc ggtttgcgta ttggccgtta acggcgggca gcgttgggtc
     1561 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     1621 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     1681 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     1741 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     1801 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     1861 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     1921 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     1981 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     2041 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     2101 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     2161 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     2221 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2281 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     2341 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     2401 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     2461 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2521 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2581 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2641 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2701 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2761 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2821 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2881 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2941 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3001 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3061 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     3121 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3181 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3241 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3301 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3361 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3421 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3481 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3541 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3601 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3661 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3721 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3781 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3841 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3901 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3961 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4021 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4081 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4141 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4201 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4261 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4321 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4381 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     4441 aaaaataggc gtatcacgag gccctttcgt cttcaagaat ccgttaacgg cattcgccat
     4501 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     4561 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     4621 cacgacgttg taaaacgacg gccagtgcca agcttggctg caggtcgacg gatccccggg
     4681 aa
//
