LOCUS       sequence-190485-        5512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Dronpa tagged ZapA in E coli.
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: pJB057
FEATURES             Location/Qualifiers
     source          1..5512
                     /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             227..901
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MVSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKV
                     KEGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNA
                     TNDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALS
                     LEGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPR
                     QAK"
     terminator      1313..1407
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1477..1499)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1964..2066
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2067..2726
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2133..2152)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3277..3865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3766..3785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4019..4036
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4109..4186
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4187..5269
                     /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(4206..4225)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5318..5323,5324..5341,5342..5348)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5356..5372
                     /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     5380..5396
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5380..5396
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5412..5429)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5412..5428)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5421..5443)
                     /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 actagtatgg tgagtgtgat
      241 taaaccagac atgaagatca agctgcgtat ggaaggcgct gtaaatggac acccgttcgc
      301 gattgaagga gttggccttg ggaagccttt cgagggaaaa cagagtatgg accttaaagt
      361 caaagaaggc ggacctctgc ctttcgccta tgacatcttg acaactgtgt tctgttacgg
      421 caacagggta ttcgccaaat acccagaaaa tatagtagac tatttcaagc agtcgtttcc
      481 tgagggctac tcttgggaac gaagcatgaa ttacgaagac gggggcattt gtaacgcgac
      541 aaacgacata accctggatg gtgactgtta tatctatgaa attcgatttg atggtgtgaa
      601 ctttcctgcc aatggtccag ttatgcagaa gaggactgtg aaatgggagc catccactga
      661 gaaattgtat gtgcgtgatg gagtgctgaa gggtgatgtt aacatggctc tgtcgcttga
      721 aggaggtggc cattaccgat gtgacttcaa aactacttat aaagctaaga aggttgtcca
      781 gttgccagac tatcactttg tggaccacca cattgagatt aaaagccacg acaaagatta
      841 cagtaatgtt aatctgcatg agcacgccga agcgcattct gagctgccga ggcaggccaa
      901 gggatccgct ggctccgctg ctggttctgg cgaattctct gcacaacccg tcgatatcca
      961 aatttttggc cgttcactgc gtgtgaactg cccgcctgac caaagggatg cgttgaatca
     1021 ggcagcggac gatctgaacc aacggttgca agatctgaaa gaacgcacta gagtcacaaa
     1081 tactgaacag ttggtcttca ttgccgcatt gaatatcagc tatgagttag cgcaagaaaa
     1141 agcaaagact cgtgactacg cggcaagtat ggaacagcgt attcggatgc tgcagcagac
     1201 catagaacaa gcgttacttg aacaaggtcg catcaccgaa aaaactaacc aaaactttga
     1261 atgagcggcc gctaagggtc gacctgcagc caagcttaat tagctgagct tggactcctg
     1321 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     1381 cgccgggcgt tttttattgg tgagaatcca agctagggga attcatcgtg actgactgac
     1441 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     1501 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     1561 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccgg tcacgtagcg atagcggagt
     1621 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1681 gataataatg gtttcttagg atccgggatg tccggcggtg cttttgccgt tacgcaccac
     1741 cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg gagcacctca
     1801 aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg
     1861 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
     1921 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag
     1981 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     2041 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     2101 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     2161 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     2221 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     2281 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     2341 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     2401 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     2461 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     2521 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     2581 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     2641 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     2701 gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc
     2761 ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat tcgaaagcaa
     2821 attcgacccg gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg tctattgctg
     2881 gtttaccggt ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg
     2941 ccagtttgct caggctctcc ccgtggaggt aataattgac gttatgatca tttattctgc
     3001 ctcccagagc ctgataaaaa cggttaccgg atcgagtcag gcaactatgg atgaacgaaa
     3061 tagacagatc gcttagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3121 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3181 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3241 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3301 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3361 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3421 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3481 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3541 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     3601 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3661 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3721 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3781 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3841 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3901 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3961 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4021 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     4081 gtgcggtatt tcacaccgca taaattccga caccatcgaa tggcgcaaaa cctttcgcgg
     4141 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     4201 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     4261 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     4321 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     4381 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     4441 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     4501 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     4561 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     4621 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     4681 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     4741 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     4801 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     4861 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     4921 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     4981 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     5041 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     5101 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     5161 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5221 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5281 ctaatgtaag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5341 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5401 attacggatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     5461 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cg
//
