LOCUS       sequence-135771-        5512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of mEos2-ZapA in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5512)
  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 5512)
  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: pJB051 (mEos2-ZapA)
FEATURES             Location/Qualifiers
     source          1..5512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      235..823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     724..743
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     977..994
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1067..1144
                     /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             1145..2227
                     /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(1164..1183)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2276..2281,2282..2299,2300..2306)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2314..2330
                     /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     2338..2354
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2338..2354
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2370..2387)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2370..2386)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2379..2401)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2586..2600,2601..2606,2607..2623,2624..2630)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    2606..2622
                     /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    2638..2654
                     /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             2673..2684
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2697..3374
                     /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      3786..3880
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3950..3972)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4436..4538
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4539..5198
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4605..4624)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
       61 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
      121 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
      181 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
      241 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
      301 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
      361 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
      421 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
      481 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
      541 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
      601 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
      661 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
      721 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
      781 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
      841 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
      901 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
      961 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     1021 ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg
     1081 gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     1141 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     1201 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     1261 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     1321 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     1381 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     1441 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     1501 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     1561 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     1621 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     1681 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     1741 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     1801 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     1861 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     1921 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
     1981 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     2041 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     2101 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     2161 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     2221 gcagtgagcg caacgcaact aatgtaagtt agctcactca ttaggcaccc caggctttac
     2281 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     2341 gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg
     2401 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2461 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcctaa gaaaccatta
     2521 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg
     2581 agaaatcata aaaaatttat ttgctttgtg agcggataac aattataata gattcaattg
     2641 tgagcggata acaatttcac acagaattca ttaaagagga gaaattaact actagtatga
     2701 gtgcgattaa gccagacatg aagatcaaac tccgtatgga aggcaacgta aacgggcacc
     2761 actttgtgat cgacggagat ggtacaggca agccttttga gggaaaacag agtatggatc
     2821 ttgaagtcaa agagggcgga cctctgcctt ttgcctttga tatcctgacc actgcattcc
     2881 attacggcaa cagggtattc gccaaatatc cagacaacat acaagactat tttaagcagt
     2941 cgtttcctaa ggggtattcg tgggaacgaa gcttgacttt cgaagacggg ggcatttgca
     3001 ttgccagaaa cgacataaca atggaagggg acactttcta taataaagtt cgattttatg
     3061 gtaccaactt tcccgccaat ggtccagtta tgcagaagaa gacgctgaaa tgggagccct
     3121 ccactgagaa aatgtatgtg cgtgatggag tgctgacggg tgatattcat atggctttgt
     3181 tgcttgaagg aaatgcccat taccgatgtg acttcagaac tacttacaaa gctaaggaga
     3241 agggtgtcaa gttaccaggc taccactttg tggaccactg cattgagatt ttaagccatg
     3301 acaaagatta caacaaggtt aagctgtatg agcatgctgt tgctcattct ggattgcctg
     3361 acaatgccag acgaggatcc gctggctccg ctgctggttc tggcgaattc tctgcacaac
     3421 ccgtcgatat ccaaattttt ggccgttcac tgcgtgtgaa ctgcccgcct gaccaaaggg
     3481 atgcgttgaa tcaggcagcg gacgatctga accaacggtt gcaagatctg aaagaacgca
     3541 ctagagtcac aaatactgaa cagttggtct tcattgccgc attgaatatc agctatgagt
     3601 tagcgcaaga aaaagcaaag actcgtgact acgcggcaag tatggaacag cgtattcgga
     3661 tgctgcagca gaccatagaa caagcgttac ttgaacaagg tcgcatcacc gaaaaaacta
     3721 accaaaactt tgaatgagcg gccgctaagg gtcgacctgc agccaagctt aattagctga
     3781 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     3841 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagctagg ggaattcatc
     3901 gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3961 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4021 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cggtcacgta
     4081 gcgatagcgg agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     4141 aggttaatgt catgataata atggtttctt aggatcctga tgtccggcgg tgcttttgcc
     4201 gttacgcacc accccgtcag tagctgaaca ggagggacag ctgatagaaa cagaagccac
     4261 tggagcacct caaaaacacc atcatacact aaatcagtaa gttggcagca tcacccgacg
     4321 cactttgcgc cgaataaata cctgtgacgg aagatcactt cgcagaataa ataaatcctg
     4381 gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga gacgttgatc
     4441 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     4501 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     4561 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     4621 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     4681 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     4741 atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg
     4801 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     4861 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     4921 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     4981 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     5041 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     5101 cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt cggcagaatg cttaatgaat
     5161 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     5221 gcccttaaac gcctggtgct acgcctgaat aagtgataat aagcggatga atggcagaaa
     5281 ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat
     5341 gtctattgct ggtttaccgg tttattgact accggaagca gtgtgaccgt gtgcttctca
     5401 aatgcctgag gccagtttgc tcaggctctc cccgtggagg taataattga cgatatgatc
     5461 atttattctg cctcccagag cctgataaaa acggttagga tcggagtcag gc
//
