LOCUS       sequence-133156-        5431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of ZapB-mEos2 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5431)
  AUTHORS   Buss J, Coltharp C, Shtengel G, Yang X, Hess H, Xiao J
  TITLE     A multi-layered protein network stabilizes the Escherichia coli 
            FtsZ-ring and modulates constriction dynamics.
  JOURNAL   PLoS Genet. 2015 Apr 7;11(4):e1005128. doi: 
            10.1371/journal.pgen.1005128. eCollection 2015 Apr.
  PUBMED    25848771
REFERENCE   2  (bases 1 to 5431)
  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: "PLoS 
            Genet."; date: "2015-04-7"; volume: "11(4):e1005128. doi"; pages: " 
            10.1371/journal.pgen.1005128. eCollection 2015 Apr"
COMMENT     Sequence Label: pJB045 (ZapB-mEos2)
FEATURES             Location/Qualifiers
     source          1..5431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(121..135,136..141,142..158,159..165)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    141..157
                     /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)."
     protein_bind    173..189
                     /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)."
     RBS             208..219
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             511..1191
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     terminator      1240..1334
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1404..1426)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1890..1992
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1993..2652
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2059..2078)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3201..3789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3690..3709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3943..3960
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4033..4110
                     /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."
     CDS             4111..5193
                     /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(4130..4149)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5242..5247,5248..5265,5266..5272)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5280..5296
                     /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     5304..5320
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5304..5320
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5336..5353)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5336..5352)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5345..5367)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cctaagaaac cattattatc
       61 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca cctcgagaaa
      121 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
      181 ggataacaat ttcacacaga attcattaaa gaggagaaat taactactag tatgacaatg
      241 tcattagaag tgtttgagaa actggaagca aaagtacagc aggcgattga taccatcact
      301 ctgttgcaga tggaaatcga agagctgaaa gaaaaaaaca actcactgtc gcaggaagtt
      361 caaaatgccc agcatcagcg cgaagagctg gagcgtgaga acaaccatct gaaagaacag
      421 cagaacggct ggcaggaacg tctgcaggcc ctgctgggtc gcatggaaga ggtcggatcc
      481 gctggctccg ctgctggttc tggcgaattc atgagtgcga ttaagccaga catgaagatc
      541 aaactccgta tggaaggcaa cgtaaacggg caccactttg tgatcgacgg agatggtaca
      601 ggcaagcctt ttgagggaaa acagagtatg gatcttgaag tcaaagaggg cggacctctg
      661 ccttttgcct ttgatatcct gaccactgca ttccattacg gcaacagggt attcgccaaa
      721 tatccagaca acatacaaga ctattttaag cagtcgtttc ctaaggggta ttcgtgggaa
      781 cgaagcttga ctttcgaaga cgggggcatt tgcattgcca gaaacgacat aacaatggaa
      841 ggggacactt tctataataa agttcgattt tatggtacca actttcccgc caatggtcca
      901 gttatgcaga agaagacgct gaaatgggag ccctccactg agaaaatgta tgtgcgtgat
      961 ggagtgctga cgggtgatat tcatatggct ttgttgcttg aaggaaatgc ccattaccga
     1021 tgtgacttca gaactactta caaagctaag gagaagggtg tcaagttacc aggctaccac
     1081 tttgtggacc actgcattga gattttaagc catgacaaag attacaacaa ggttaagctg
     1141 tatgagcatg ctgttgctca ttctggattg cctgacaatg ccagacgata agcggccgct
     1201 aagggtcgac ctgcagccaa gcttaattag ctgagcttgg actcctgttg atagatccag
     1261 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     1321 ttattggtga gaatccaagc taggggaatt catcgtgact gactgacgat ctgcctcgcg
     1381 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     1441 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     1501 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga
     1561 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1621 tcttaggatc ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcagtagctg
     1681 aacaggaggg acagctgata gaaacagaag ccactggagc acctcaaaaa caccatcata
     1741 cactaaatca gtaagttggc agcatcaccc gacgcacttt gcgccgaata aatacctgtg
     1801 acggaagatc acttcgcaga ataaataaat cctggtgtcc ctgttgatac cgggaagccc
     1861 tgggccaact tttggcgaaa atgagacgtt gatcggcacg taagaggttc caactttcac
     1921 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     1981 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
     2041 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     2101 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     2161 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca
     2221 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
     2281 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     2341 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     2401 gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt
     2461 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     2521 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtctgt
     2581 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     2641 ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg tgctacgcct
     2701 gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg acccggtcgt
     2761 cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta ccggtttatt
     2821 gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt ttgctcaggc
     2881 tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc agagcctgat
     2941 aaaaacggtt aggatcggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3001 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3061 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3121 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3181 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3241 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3301 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3361 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3421 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3481 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3541 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3601 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3661 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3721 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3781 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3841 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3901 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3961 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4021 cgcataaatt ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc
     4081 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
     4141 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     4201 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     4261 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     4321 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     4381 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     4441 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     4501 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     4561 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
     4621 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     4681 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     4741 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     4801 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     4861 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
     4921 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     4981 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     5041 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     5101 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     5161 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct
     5221 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     5281 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg
     5341 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     5401 cagcacatcc ccctttcgcc agctggcgta a
//
