LOCUS       sequence-194066-        7460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Dronpa tagged ZapA and pAmCherry1 tagged FtsZ in E coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7460)
  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 7460)
  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: pJB089
FEATURES             Location/Qualifiers
     source          1..7460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      22..116
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(186..208)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        673..775
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             776..1435
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(842..861)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1986..2574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2475..2494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2728..2745
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2818..2895
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2896..3978
                     /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(2915..2934)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4027..4032,4033..4050,4051..4057)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4065..4081
                     /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     4089..4105
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4089..4105
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4121..4138)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4121..4137)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4130..4152)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(4337..4351,4352..4357,4358..4374,4375..4381)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    4357..4373
                     /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    4389..4405
                     /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             4424..4435
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4448..5122
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MVSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKV
                     KEGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNA
                     TNDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALS
                     LEGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPR
                     QAK"
     CDS             6723..7433
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein"
                     /label=PAmCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6868..6886)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7136..7155
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
       61 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
      121 gctaggggaa ttcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt
      181 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
      241 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
      301 atgacccggt cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg
      361 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagga tccgggatgt
      421 ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag ctgaacagga gggacagctg
      481 atagaaacag aagccactgg agcacctcaa aaacaccatc atacactaaa tcagtaagtt
      541 ggcagcatca cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc
      601 agaataaata aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg
      661 aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
      721 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
      781 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
      841 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
      901 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
      961 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     1021 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     1081 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     1141 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     1201 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     1261 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     1321 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     1381 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt
     1441 ttaaggcagt tattggtgcc cttaaacgcc tggttgctac gcctgaataa gtgataataa
     1501 gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt
     1561 cgttaaatag ccgcttatgt ctattgctgg tttaccggtt tattgactac cggaagcagt
     1621 gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta
     1681 ataattgacg ttatgatcat ttattctgcc tcccagagcc tgataaaaac ggttaccgga
     1741 tcgagtcagg caactatgga tgaacgaaat agacagatcg cttagatagg tgcctcactg
     1801 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1861 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1921 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1981 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2041 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2101 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2161 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2221 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2281 gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2341 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2401 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2461 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2521 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2581 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2641 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2701 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2761 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac
     2821 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     2881 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     2941 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     3001 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     3061 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     3121 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     3181 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     3241 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     3301 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     3361 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     3421 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     3481 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     3541 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     3601 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     3661 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     3721 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     3781 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     3841 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     3901 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     3961 ctggaaagcg ggcagtgagc gcaacgcaac taatgtaagt tagctcactc attaggcacc
     4021 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     4081 atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac
     4141 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4201 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgccta
     4261 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4321 tcttcacctc gagaaatcat aaaaaattta tttgctttgt gagcggataa caattataat
     4381 agattcaatt gtgagcggat aacaatttca cacagaattc attaaagagg agaaattaac
     4441 tactagtatg gtgagtgtga ttaaaccaga catgaagatc aagctgcgta tggaaggcgc
     4501 tgtaaatgga cacccgttcg cgattgaagg agttggcctt gggaagcctt tcgagggaaa
     4561 acagagtatg gaccttaaag tcaaagaagg cggacctctg cctttcgcct atgacatctt
     4621 gacaactgtg ttctgttacg gcaacagggt attcgccaaa tacccagaaa atatagtaga
     4681 ctatttcaag cagtcgtttc ctgagggcta ctcttgggaa cgaagcatga attacgaaga
     4741 cgggggcatt tgtaacgcga caaacgacat aaccctggat ggtgactgtt atatctatga
     4801 aattcgattt gatggtgtga actttcctgc caatggtcca gttatgcaga agaggactgt
     4861 gaaatgggag ccatccactg agaaattgta tgtgcgtgat ggagtgctga agggtgatgt
     4921 taacatggct ctgtcgcttg aaggaggtgg ccattaccga tgtgacttca aaactactta
     4981 taaagctaag aaggttgtcc agttgccaga ctatcacttt gtggaccacc acattgagat
     5041 taaaagccac gacaaagatt acagtaatgt taatctgcat gagcacgccg aagcgcattc
     5101 tgagctgccg aggcaggcca agggatccgc tggctccgct gctggttctg gcgaattctc
     5161 tgcacaaccc gtcgatatcc aaatttttgg ccgttcactg cgtgtgaact gcccgcctga
     5221 ccaaagggat gcgttgaatc aggcagcgga cgatctgaac caacggttgc aagatctgaa
     5281 agaacgcact agagtcacaa atactgaaca gttggtcttc attgccgcat tgaatatcag
     5341 ctatgagtta gcgcaagaaa aagcaaagac tcgtgactac gcggcaagta tggaacagcg
     5401 tattcggatg ctgcagcaga ccatagaaca agcgttactt gaacaaggtc gcatcaccga
     5461 aaaaactaac caaaactttg aatgagtcga cctgcaggca tgcaagcttt aactttaaga
     5521 aggagatata cgctagcatg tttgaaccaa tggaacttac caatgacgcg gtgattaaag
     5581 tcatcggcgt cggcggcggc ggcggtaatg ctgttgaaca catggtgcgc gagcgcattg
     5641 aaggtgttga attcttcgcg gtaaataccg atgcacaagc gctgcgtaaa acagcggttg
     5701 gacagacgat tcaaatcggt agcggtatca ccaaaggact gggcgctggc gctaatccag
     5761 aagttggccg caatgcggct gatgaggatc gcgatgcatt gcgtgcggcg ctggaaggtg
     5821 cagacatggt ctttattgct gcgggtatgg gtggtggtac cggtacaggt gcagcaccag
     5881 tcgtcgctga agtggcaaaa gatttgggta tcctgaccgt tgctgtcgtc actaagcctt
     5941 tcaactttga aggcaagaag cgtatggcat tcgcggagca ggggatcact gaactgtcca
     6001 agcatgtgga ctctctgatc actatcccga acgacaaact gctgaaagtt ctgggccgcg
     6061 gtatctccct gctggatgcg tttggcgcag cgaacgatgt actgaaaggc gctgtgcaag
     6121 gtatcgctga actgattact cgtccgggtt tgatgaacgt ggactttgca gacgtacgca
     6181 ccgtaatgtc tgagatgggc tacgcaatga tgggttctgg cgtggcgagc ggtgaagacc
     6241 gtgcggaaga agctgctgaa atggctatct cttctccgct gctggaagat atcgacctgt
     6301 ctggcgcgcg cggcgtgctg gttaacatca cggcgggctt cgatctgcgt ctggatgagt
     6361 tcgaaacggt aggtaacacc atccgtgcat ttgcttccga caacgcgact gtggttatcg
     6421 gtacttctct tgacccggat atgaatgacg agctgcgcgt aaccgttgtt gcgacaggta
     6481 tcggcatgga caaacgtcct gaaatcactc tggtgaccaa taagcaggtt cagcagccag
     6541 tgatggatcg ctaccagcag catgggatgg ctccgctgac ccaggagcag aagccggttg
     6601 ctaaagtcgt gaatgacaat gcgccgcaaa ctgcgaaaga gccggattat ctggatatcc
     6661 cagcattcct gcgtaagcaa gctgatggat ccgctggctc cgctgctggt tctggcgaat
     6721 tcatggtgag caagggcgag gaggataaca tggccatcat taaggagttc atgcgcttca
     6781 aggtgcacat ggaggggtcc gtgaacggcc acgtgttcga gatcgagggc gagggcgagg
     6841 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     6901 ccttcacctg ggacatcctg tcccctcaat tcatgtacgg ctccaatgcc tacgtgaagc
     6961 accccgccga catccccgac tactttaagc tgtccttccc cgagggcttc aagtgggagc
     7021 gcgtgatgaa attcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     7081 acggtgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     7141 taatgcagaa gaagaccatg ggctgggagg ccctctccga gcggatgtac cccgaggacg
     7201 gcgccctgaa gggcgaggtc aagccgagag tgaagctgaa ggacggcggc cactacgacg
     7261 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     7321 tcaaccgcaa gttggacatc acctcacaca acgaggacta caccatcgtg gaacagtacg
     7381 aacgtgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taagcggccg
     7441 ctaagggtcg acctgcagcc
//
