LOCUS       sequence-177657-        4700 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Constitutive expression of a chimeric LacI:PurR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4700)
  AUTHORS   Tungtur S, Egan SM, Swint-Kruse L
  TITLE     Functional consequences of exchanging domains between LacI and PurR 
            are mediated by the intervening linker sequence.
  JOURNAL   Proteins. 2007 Jul 1;68(1):375-88.
  PUBMED    17436321
REFERENCE   2  (bases 1 to 4700)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proteins.";
            date: "2007-07-1"; volume: "68(1)"; pages: "375-88"
COMMENT     Sequence Label: LLhP-pHG165
FEATURES             Location/Qualifiers
     source          1..4700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..27
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11..27
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        68..145
                     /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(165..184)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1400..1416)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1400..1416)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1413..1435)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1424..1440
                     /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(1448..1454,1455..1472,1473..1478))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1493..1514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2114..2305
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2299..2321)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2407..2547
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2462..2481
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2562..2579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2733..3321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2813..2832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3492..4283,4284..4352))
                     /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     4115..4134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4353..4457)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4525..4543
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     join(4696..4700,1..18)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tcacgacgtt gtaaaacgac ggccagtgcc aagcttggct gcaggtcgac ggatccccgg
       61 gaattccgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga
      121 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
      181 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
      241 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctc aattacattc ccaaccgcgt
      301 ggcacaacaa ctggcgggca aacagagcaa atctatcggt ttgctggcga ccagcagcga
      361 agcggcctat tttgccgaga tcattgaagc agttgaaaaa aattgcttcc agaaaggtta
      421 caccctgatt ctgggcaatg cgtggaacaa tcttgagaaa cagcgggctt atctgtcgat
      481 gatggcgcaa aaacgcgtcg atggtctgct ggtgatgtgt tctgagtacc cagagccgtt
      541 gctggcgatg ctggaagagt atcgccatat cccaatggtg gtcatggact ggggtgaagc
      601 aaaagctgac ttcaccgatg cggtcattga taacgcgttc gaaggcggct acatggccgg
      661 gcgttatctg attgaacgcg gtcaccgcga aatcggcgtc atccccggcc cgctggaacg
      721 taacaccggc gcaggccgcc ttgccggttt tatgaaggcg atggaagaag cgatgatcaa
      781 ggtgccggaa agctggattg tgcagggtga ctttgaacct gaatccggtt atcgcgccat
      841 gcagcaaatc ctgtcgcagc cgcatcgccc tactgccgtc ttctgtggtg gcgatatcat
      901 ggcaatgggc gcactttgtg ctgctgatga aatgggcctg cgcgtcccgc aggatgtttc
      961 gctgatcggt tatgataacg tgcgcaacgc gcgctatttt acgccggcgc tgaccacgat
     1021 ccatcagcca aaagattcgc tgggtgaaac agcgttcaac atgctgttgg atcgtatcgt
     1081 caacaaacgt gaagaaccgc agtctattga agtgcatccg cgcttgattg aacgccgctc
     1141 cgtggctgac ggcccgttcc gcgactatcg tcgttaataa cctgaggccg atactgtcgt
     1201 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta
     1261 tcccattagg tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc
     1321 acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc
     1381 gttggaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1441 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1501 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1561 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attggccgtt
     1621 aacggcgggc agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag
     1681 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga
     1741 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
     1801 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
     1861 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
     1921 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
     1981 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
     2041 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
     2101 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
     2161 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
     2221 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
     2281 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2341 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2401 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     2461 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     2521 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     2581 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2641 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2701 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2761 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2821 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2881 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2941 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3001 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3061 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3121 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     3181 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3241 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3301 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3361 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3421 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3481 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3541 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3601 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3661 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3721 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3781 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3841 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3901 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3961 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4021 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4081 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4141 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4201 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4261 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4321 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4381 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4441 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4501 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
     4561 tccgttaacg gcattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4621 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4681 taacgccagg gttttcccag
//
