LOCUS       sequence-190376-        5410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Dronpa tagged ZapB in E coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5410)
  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 5410)
  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: pJB073
FEATURES             Location/Qualifiers
     source          1..5410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(116..130,131..136,137..153,154..160)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    136..152
                     /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    168..184
                     /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             203..214
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             485..1162
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MVSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKV
                     KEGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNA
                     TNDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALS
                     LEGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPR
                     QAK"
     terminator      1211..1305
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1375..1397)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1862..1964
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1965..2624
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2031..2050)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3175..3763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3664..3683
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3917..3934
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4007..4084
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4085..5167
                     /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(4104..4123)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5216..5221,5222..5239,5240..5246)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5254..5270
                     /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     5278..5294
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5278..5294
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5310..5327)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5310..5326)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5319..5341)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aagaggcccg caccgatcgc ccttcccaac agttgcctaa gaaaccatta ttatcatgac
       61 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg agaaatcata
      121 aaaaatttat ttgctttgtg agcggataac aattataata gattcaattg tgagcggata
      181 acaatttcac acagaattca ttaaagagga gaaattaact actagtatga caatgtcatt
      241 agaagtgttt gagaaactgg aagcaaaagt acagcaggcg attgatacca tcactctgtt
      301 gcagatggaa atcgaagagc tgaaagaaaa aaacaactca ctgtcgcagg aagttcaaaa
      361 tgcccagcat cagcgcgaag agctggagcg tgagaacaac catctgaaag aacagcagaa
      421 cggctggcag gaacgtctgc aggccctgct gggtcgcatg gaagaggtca gcatccccgg
      481 cgccatggtg agtgtgatta aaccagacat gaagatcaag ctgcgtatgg aaggcgctgt
      541 aaatggacac ccgttcgcga ttgaaggagt tggccttggg aagcctttcg agggaaaaca
      601 gagtatggac cttaaagtca aagaaggcgg acctctgcct ttcgcctatg acatcttgac
      661 aactgtgttc tgttacggca acagggtatt cgccaaatac ccagaaaata tagtagacta
      721 tttcaagcag tcgtttcctg agggctactc ttgggaacga agcatgaatt acgaagacgg
      781 gggcatttgt aacgcgacaa acgacataac cctggatggt gactgttata tctatgaaat
      841 tcgatttgat ggtgtgaact ttcctgccaa tggtccagtt atgcagaaga ggactgtgaa
      901 atgggagcca tccactgaga aattgtatgt gcgtgatgga gtgctgaagg gtgatgttaa
      961 catggctctg tcgcttgaag gaggtggcca ttaccgatgt gacttcaaaa ctacttataa
     1021 agctaagaag gttgtccagt tgccagacta tcactttgtg gaccaccaca ttgagattaa
     1081 aagccacgac aaagattaca gtaatgttaa tctgcatgag cacgccgaag cgcattctga
     1141 gctgccgagg caggccaagt aagcggccgc taagggtcga cctgcagcca agcttaatta
     1201 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     1261 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag ctaggggaat
     1321 tcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     1381 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1441 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccggtc
     1501 acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt
     1561 tttataggtt aatgtcatga taataatggt ttcttaggat ccgggatgtc cggcggtgct
     1621 tttgccgtta cgcaccaccc cgtcagtagc tgaacaggag ggacagctga tagaaacaga
     1681 agccactgga gcacctcaaa aacaccatca tacactaaat cagtaagttg gcagcatcac
     1741 ccgacgcact ttgcgccgaa taaatacctg tgacggaaga tcacttcgca gaataaataa
     1801 atcctggtgt ccctgttgat accgggaagc cctgggccaa cttttggcga aaatgagacg
     1861 ttgatcggca cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg
     1921 tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
     1981 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     2041 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
     2101 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     2161 tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg
     2221 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     2281 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     2341 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     2401 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     2461 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     2521 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     2581 atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt
     2641 attggtgccc ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg
     2701 gcagaaattc gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc
     2761 cgcttatgtc tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg
     2821 cttctcaaat gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacgt
     2881 tatgatcatt tattctgcct cccagagcct gataaaaacg gttaccggat cgagtcaggc
     2941 aactatggat gaacgaaata gacagatcgc ttagataggt gcctcactga ttaagcattg
     3001 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3061 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3121 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3181 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3241 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3301 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3361 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3421 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3481 gcggtcggac tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3541 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3601 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3661 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3721 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3781 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3841 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3901 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     3961 ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg
     4021 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     4081 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     4141 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     4201 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     4261 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     4321 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     4381 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     4441 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     4501 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     4561 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     4621 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     4681 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     4741 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     4801 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     4861 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
     4921 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     4981 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     5041 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     5101 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5161 gcagtgagcg caacgcaact aatgtaagtt agctcactca ttaggcaccc caggctttac
     5221 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5281 gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg
     5341 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     5401 cgtaatagcg
//
