LOCUS       sequence-178284-        4704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:GalR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4704)
  AUTHORS   Tungtur S, Meinhardt S, Swint-Kruse L
  TITLE     Comparing the functional roles of nonconserved sequence positions in
            homologous transcription repressors: implications for 
            sequence/function analyses.
  JOURNAL   J Mol Biol. 2010 Jan 29;395(4):785-802. doi: 
            10.1016/j.jmb.2009.10.001. Epub 2009 Oct 8.
  PUBMED    19818797
REFERENCE   2  (bases 1 to 4704)
  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: 
            "10.1016/j.jmb.2009.10.001"; journalName: "J Mol Biol"; date: 
            "2010-01-29- 29"; volume: "395"; issue: "4"; pages: "785-802"
COMMENT     Sequence Label: LGhG-pHG165a
FEATURES             Location/Qualifiers
     source          1..4704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..89
                     /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(109..128)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1374..1391)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1374..1390)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1383..1405)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1558..1576)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1644..1748
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1749..1817,1818..2609)
                     /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     complement(1967..1986)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2780..3368
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3269..3288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3522..3539
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3554..3694
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3620..3639)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3780..3802
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3796..3987)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    4587..4608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4623..4628,4629..4646,4647..4653)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     4683..4699
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4683..4699
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttacgaattc cgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc
       61 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
      121 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
      181 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctcagctat cacccgaacg
      241 ccaacgcccg tgcgctggcg cagcagacca ctgaaacggt cggtctggtc gttggtgatg
      301 tttccgatcc gtttttcggt gcaatggtga aagcggtcga acaggtggct tatcacaccg
      361 gtaatttttt attgattggc aacggttacc acaacgaaca aaaagagcgt caggccattg
      421 agcaactgat ccgccatcgc tgtgctgcgt tggtcgtcca tgccaaaatg atcccggatg
      481 ctgatttagc ctcattaatg aaacaaatgc ccggtatggt gctgatcaac cgtatcctgc
      541 ctggctttga aaaccgttgt attgctctgg acgatcgtta cggtgcctgg ctggcaacgc
      601 gtcatttaat tcagcaaggt catacccgca ttggttatct gtgctctaac cactctattt
      661 ctgacgccga agatcgtctg caagggtatt acgatgccct tgctgaaagt ggtattgcgg
      721 ccaatgaccg gctggtgaca tttggcgaac cagacgaaag cggcggcgaa caggcaatga
      781 ccaagctttt gggacgagga agaaatttca ctgcggtagc ctgttataac gattcaatgg
      841 cggcgggtgc gatgggcgtt ctcaatgata atggtattga tgtaccgggt gagatttcgt
      901 taattggctt tgatgatgtg ctggtgtcac gctatgtgcg tccgcgcctg accaccgtgc
      961 gttacccaat cgtgacgatg gcgacccagg ctgccgaact ggctttggcg ctggcggata
     1021 atcgccctct cccggaaatc actaatgtct ttagtccgac gctggtacgt cgtcattcag
     1081 tgtcaactcc gtcgctggag gcaagtcatc atgcaaccag cgactaataa cctgaggccg
     1141 atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca
     1201 acgtgaccta tcccattagg tcaatccgcc gtttgttccc acggagaatc cgacgggttg
     1261 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat
     1321 ttttgatggc gttggaattc ccggggatcc gtcgacctgc agccaagctt ggcactggcc
     1381 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     1441 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1501 caacagttgc gcagcctgaa tggcgaatgc cgttaacgga ttcttgaaga cgaaagggcc
     1561 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     1621 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1681 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1741 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1801 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1861 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1921 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1981 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2041 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2101 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2161 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2221 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2281 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2341 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2401 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2461 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2521 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2581 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2641 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2701 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2761 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2821 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2881 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     2941 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3001 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3061 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3121 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3181 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3241 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3301 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3361 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3421 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3481 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3541 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3601 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3661 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3721 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3781 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3841 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3901 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3961 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     4021 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     4081 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     4141 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     4201 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     4261 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     4321 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     4381 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     4441 catgcgcacc cgtggccagg acccaacgct gcccgccgtt aacggccaat acgcaaaccg
     4501 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4561 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4621 gctttacact ttatgcttcc ggctcgtatg ttgtgtggcg tggtactcat aacaatttca
     4681 cacaggaaac agctatgacc atga
//
