LOCUS       sequence-177280-        3822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty control plasmid, three terminators to place inserts between 
            for insulation, ampR, pBR322 origin, lacI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3822)
  AUTHORS   Kelly CL, Liu Z, Yoshihara A, Jenkinson SF, Wormald MR, Otero J, 
            Estevez A, Kato A, Marqvorsen MH, Fleet GW, Estevez RJ, Izumori K, 
            Heap JT
  TITLE     Synthetic Chemical Inducers and Genetic Decoupling Enable Orthogonal
            Control of the rhaBAD Promoter.
  JOURNAL   ACS Synth Biol. 2016 Oct 21;5(10):1136-1145. Epub 2016 Jun 21.
  PUBMED    27247275
REFERENCE   2  (bases 1 to 3822)
  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: "ACS Synth 
            Biol."; date: "2016-10-21"; volume: "5(10)"; pages: "1136-1145. Epub
            2016 Jun 21"
COMMENT     Sequence Label: pCK300
FEATURES             Location/Qualifiers
     source          1..3822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      86..674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     575..594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     828..845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     943..960
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     944..960
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1619..1638)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1644..1663)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        1705..1796
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1797..1865,1866..2657)
                     /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(2015..2034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             2740..3822
                     /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="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2759..2778)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc
       61 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      241 tgtagccgta gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      841 gcgaggaagc ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc
      901 tgacggatgg cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt
      961 cttaagctcg ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg
     1021 taaaaacccg cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag
     1081 aatatttaag ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg
     1141 agaaaaaagc aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata
     1201 attaaactat tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa
     1261 agagaattaa gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa
     1321 attatacatg tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt
     1381 attatgcatt tagaataaat tttgtgtcgc cctttaactc gagccccaag ggcgacaccc
     1441 cataattagc ccgggcgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc
     1501 ctggcagttc cctactctcg catggggagt ccccacacta ccatcggcgc tacggcgttt
     1561 cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaaca
     1621 aggggtgtta tgagccatat tcaggtataa atgggctcgc gataatgttc agaattggtt
     1681 aattggttgt aacactgacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1741 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagaatatga
     1801 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1861 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1921 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1981 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2041 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2101 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2161 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2221 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2281 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2341 tagcgatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2401 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2461 cccttccggc tggctggttt attgctgata aatccggagc cggtgagcgt ggttctcgcg
     2521 gtatcatcgc agcgctgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2581 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2641 tgattaagca ttggtaagcg gcgcgccatc gaatggcgca aaacctttcg cggtatggca
     2701 tgatagcgcc cggaagagag tcaattcagg gtggtgaata tgaaaccagt aacgttatac
     2761 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     2821 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     2881 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     2941 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     3001 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     3061 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     3121 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     3181 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga ggacggtacg
     3241 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     3301 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     3361 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     3421 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     3481 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     3541 atctcggtag tgggatacga cgataccgaa gatagctcat gttatatccc gccgttaacc
     3601 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     3661 tctcagggcc aggcggtgaa gggcaatcag ctgttgccag tctcactggt gaaaagaaaa
     3721 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3781 cagctggcac gacaggtttc ccgactggaa agcgggcagt ga
//
