LOCUS       sequence-178201-        4676 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Constitutive expression of a chimeric LacI:CytR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4676)
  AUTHORS   Meinhardt S, Manley MW Jr, Becker NA, Hessman JA, Maher LJ 3rd, 
            Swint-Kruse L
  TITLE     Novel insights from hybrid LacI/GalR proteins: family-wide 
            functional attributes and biologically significant variation in 
            transcription repression.
  JOURNAL   Nucleic Acids Res. 2012 Nov;40(21):11139-54. doi: 
            10.1093/nar/gks806. Epub 2012 Sep 10.
  PUBMED    22965134
REFERENCE   2  (bases 1 to 4676)
  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"; doi: 
            "10.1093/nar/gks806"; journalName: "Nucleic Acids Res"; date: 
            "2012-11"; volume: "40"; issue: "21"; pages: "11139-54"
COMMENT     Sequence Label: LLhC-pHG165a
FEATURES             Location/Qualifiers
     source          1..4676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(149..217,218..1009)
                     /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(367..386)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1180..1768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1669..1688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1922..1939
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1954..2094
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2020..2039)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2180..2202
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2196..2387)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    2987..3008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3023..3028,3029..3046,3047..3053)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3061..3077
                     /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)."
     primer_bind     3066..3088
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3085..3101
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3085..3101
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3118..3195
                     /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(3215..3234)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(4450..4467)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4450..4466)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4459..4481)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4634..4652)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
       61 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      121 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      181 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      241 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      301 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      361 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
      421 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      481 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      541 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      601 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      661 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
      721 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      781 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      841 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      901 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
      961 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1021 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1081 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1141 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1201 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1261 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1321 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1381 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1441 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1501 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1561 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1621 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1681 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1741 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1801 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1861 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1921 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     1981 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     2041 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     2101 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     2161 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     2221 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     2281 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     2341 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     2401 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     2461 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     2521 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     2581 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     2641 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     2701 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     2761 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     2821 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgccgtt
     2881 aacggccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     2941 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     3001 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     3061 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gaattccgac
     3121 accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     3181 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     3241 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     3301 gaaaaagtgg aagcggcgat ggcggagctc aattacattc ccaaccgcgt ggcacaacaa
     3361 ctggcgggca aacagagccg caccattctg gtgattgtcc cggatatctg cgatcccttc
     3421 tttagcgaaa ttattcgcgg tatcgaagtt acggcggcaa atcacggata tctggtgctg
     3481 attggcgact gtgcgcatca aaatcagcag gaaaaaacct ttatcgattt gatcatcacc
     3541 aagcaaattg atggcatgtt gttgctgggt tcaaggctgc cgtttgatgc cagcattgag
     3601 gaacagcgta atctgccgcc gatggtgatg gcgaacgaat ttgcaccgga gctggagctg
     3661 cctacagttc atatcgacaa tctgaccgcc gcatttgatg cagtaaatta tttatatgag
     3721 caagggcata aacggattgg ctgtatagcc ggtcccgaag agatgccgct gtgtcactac
     3781 cgcctgcaag gctatgttca ggcgctgcgt cgctgcggca ttatggttga tccgcaatac
     3841 atcgcccgtg gcgacttcac cttcgaagcc ggaagcaaag cgatgcagca gctgcttgat
     3901 ctcccacaac cgcctactgc tgtcttctgc catagcgatg tgatggcgct cggcgcactt
     3961 tctcaggcaa aacgccaggg gctgaaagtc ccggaagacc tttccataat cggttttgat
     4021 aacatcgacc tgacgcaatt ttgtgatccg ccgctgacaa ccatcgcgca gccgcgttac
     4081 gaaatcggtc gggaagctat gctgttattg cttgatcaaa tgcaggggca acacgttggc
     4141 agtggctctc gtttaatgga ctgcgaactt atcatccggg gatcaacacg cgcgttacct
     4201 taataacctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat
     4261 gcgcccatct acaccaacgt gacctatccc attaggtcaa tccgccgttt gttcccacgg
     4321 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     4381 gccagacgcg aattattttt gatggcgttg gaattcccgg ggatccgtcg acctgcagcc
     4441 aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     4501 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     4561 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgccgtt aacggattct
     4621 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataat
//
