LOCUS       sequence-190310-        6996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FtsI tagged with tagRFP-t in E coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6996)
  AUTHORS   Yang X, Lyu Z, Miguel A, McQuillen R, Huang KC, Xiao J
  TITLE     GTPase activity-coupled treadmilling of the bacterial tubulin FtsZ 
            organizes septal cell wall synthesis.
  JOURNAL   Science. 2017 Feb 17;355(6326):744-747. doi: 
            10.1126/science.aak9995.
  PUBMED    28209899
REFERENCE   2  (bases 1 to 6996)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2017-02-17- 17"; volume: "355"; 
            issue: "6326"; pages: "744-747"
COMMENT     Sequence Label: pJB007
FEATURES             Location/Qualifiers
     source          1..6996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(80..94,95..100,101..117,118..124)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    100..116
                     /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    132..148
                     /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             167..178
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             191..922
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     terminator      2761..2855
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2925..2947)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3412..3514
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3515..4174
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3581..3600)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      4725..5313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5214..5233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5467..5484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5557..5634
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5635..6717
                     /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(5654..5673)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6766..6771,6772..6789,6790..6796)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6804..6820
                     /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     6828..6844
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6828..6844
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6860..6877)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6860..6876)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6869..6891)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
       61 tcgtcttcac ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat
      121 aatagattca attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt
      181 aactactagt atggtgtcta agggcgaaga gctgattaag gagaacatgc acatgaagct
      241 gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa
      301 gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt
      361 cgccttcgac atcctggcta ccagcttcat gtacggcagc agaaccttca tcaaccacac
      421 ccagggcatc cccgatttct ttaagcagtc cttccctgag ggcttcacat gggagagagt
      481 caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg
      541 ctgcctcatc tacaacgtca agatcagagg ggtgaacttc ccatccaacg gccctgtgat
      601 gcagaagaaa acactcggct gggaggccaa caccgagatg ctgtaccccg ctgacggcgg
      661 cctggaaggc agaaccgaca tggccctgaa gctcgtgggc gggggccacc tgatctgcaa
      721 cttcaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta
      781 ctatgtggac cacagactgg aaagaatcaa ggaggccgac aaagagacct acgtcgagca
      841 gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc acaaacttaa
      901 tggcatggac gagctgtaca agggatccgc tggctccgct gctggttctg gcgaattcat
      961 gaaagcagcg gcgaaaacgc agaaaccaaa acgtcaggaa gaacatgcca actttatcag
     1021 ttggcgtttt gcgttgttat gcggctgtat tctcctggcg ctggcttttc tgctcggacg
     1081 cgtagcgtgg ttacaagtta tctccccgga tatgctggtg aaagagggcg acatgcgttc
     1141 tcttcgcgtt cagcaagttt ccacctcccg cggcatgatt actgaccgtt ctggtcgccc
     1201 gttagcggtg agcgtgccgg taaaagcgat ttgggctgac ccgaaagaag tgcatgacgc
     1261 tggcggtatc agcgtcggtg accgctggaa ggcgctggct aacgcgctca atattccgct
     1321 ggatcagctt tcagcccgca ttaacgccaa cccgaaaggg cgctttattt atctggcgcg
     1381 tcaggtgaac cctgacatgg cggactacat caaaaaactg aaactgccgg ggattcatct
     1441 gcgtgaagaa tctcgccgtt attatccatc cggcgaagtg actgctcacc tcattggctt
     1501 tactaacgtc gatagtcaag ggattgaggg cgttgagaag agtttcgata aatggcttac
     1561 cgggcagccg ggtgagcgca ttgtgcgtaa agaccgctat ggtcgcgtaa ttgaagatat
     1621 ttcttctact gacagccagg ctgcgcacaa cctggcgctg agtattgatg aacgcctgca
     1681 ggcgctggtt tatcgcgaac tgaacaatgc ggtggccttt aacaaggctg aatctggtag
     1741 cgccgtgctg gtggatgtca acaccggtga agtgctggcg atggctaaca gcccgtcata
     1801 caaccctaac aatctgagcg gcacgccgaa agaggcgatg cgtaaccgta ccatcaccga
     1861 cgtgtttgaa cctggctcaa cggttaaacc gatggtggtg atgaccgcgt tgcaacgtgg
     1921 cgtggtgcgg gaaaactcgg tactcaatac cattccttat cgaattaacg gccacgaaat
     1981 caaagacgtg gcacgctaca gcgaattaac cctgaccggg gtattacaga agtcgagtaa
     2041 cgtcggtgtt tccaagctgg cgttagcgat gccgtcctca gcgttagtag atacttactc
     2101 acgttttgga ctgggaaaag cgaccaattt ggggttggtc ggagaacgca gtggcttata
     2161 tcctcaaaaa caacggtggt ctgacataga gagggccacc ttctctttcg gctacgggct
     2221 aatggtaaca ccgttacagt tagcgcgagt ctacgcaact atcggcagct acggcattta
     2281 tcgcccactg tcgattacca aagttgaccc cccggttccc ggtgaacgtg tcttcccgga
     2341 atccattgtc cgcaccgtgg tgcatatgat ggaaagcgtg gcgctaccgg gcggcggcgg
     2401 cgtgaaggcg gcgattaaag gctatcgtat cgccattaaa accggtaccg cgaaaaaggt
     2461 cgggccggac ggtcgctaca tcaataaata tattgcttat accgcaggcg ttgcgcctgc
     2521 gagtcagccg cgcttcgcgc tggttgttgt tatcaacgat ccgcaggcgg gtaaatacta
     2581 cggcggcgcc gtttccgcgc cggtctttgg tgccatcatg ggcggcgtat tgcgtaccat
     2641 gaacatcgag ccggatgcgc tgacaacggg cgataaaaat gaatttgtga ttaatcaagg
     2701 cgaggggaca ggtggcagat cgtaagtcga cctgcagcca agcttaatta gctgagcttg
     2761 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
     2821 tcggttgccg ccgggcgttt tttattggtg agaatccaag ctaggggaat tcatcgtgac
     2881 tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2941 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     3001 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccggtc acgtagcgat
     3061 agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3121 aatgtcatga taataatggt ttcttaggat ccgggatgtc cggcggtgct tttgccgtta
     3181 cgcaccaccc cgtcagtagc tgaacaggag ggacagctga tagaaacaga agccactgga
     3241 gcacctcaaa aacaccatca tacactaaat cagtaagttg gcagcatcac ccgacgcact
     3301 ttgcgccgaa taaatacctg tgacggaaga tcacttcgca gaataaataa atcctggtgt
     3361 ccctgttgat accgggaagc cctgggccaa cttttggcga aaatgagacg ttgatcggca
     3421 cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg tattttttga
     3481 gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     3541 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     3601 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     3661 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     3721 tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     3781 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     3841 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     3901 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     3961 ctgggtgagt ttcaccagtt ttgatttaaa cgtggctaat atggacaact tcttcgcccc
     4021 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     4081 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     4141 acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc
     4201 ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg gcagaaattc
     4261 gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc
     4321 tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat
     4381 gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacgt tatgatcatt
     4441 tattctgcct cccagagcct gataaaaacg gttaccggat cgagtcaggc aactatggat
     4501 gaacgaaata gacagatcgc ttagataggt gcctcactga ttaagcattg gtaactgtca
     4561 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4621 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4681 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4741 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4801 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4861 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4921 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4981 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac
     5041 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5101 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5161 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5221 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5281 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5341 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5401 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5461 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     5521 acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg gcgcaaaacc
     5581 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     5641 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     5701 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     5761 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     5821 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     5881 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     5941 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     6001 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     6061 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     6121 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     6181 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     6241 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     6301 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     6361 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     6421 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     6481 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     6541 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     6601 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6661 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     6721 caacgcaact aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct
     6781 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     6841 tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     6901 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     6961 aagaggcccg caccgatcgc ccttcccaac agttgc
//
