LOCUS       sequence-418289-        6696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing FtsW-TagRFP-T in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6696)
  AUTHORS   Yang X, McQuillen R, Lyu Z, Phillips-Mason P, De La Cruz A, 
            McCausland JW, Liang H, DeMeester KE, Santiago CC, Grimes CL, de 
            Boer P, Xiao J
  TITLE     A two-track model for the spatiotemporal coordination of bacterial 
            septal cell wall synthesis revealed by single-molecule imaging of 
            FtsW.
  JOURNAL   Nat Microbiol. 2021 May;6(5):584-593. doi: 
            10.1038/s41564-020-00853-0. Epub 2021 Jan 25.
  PUBMED    33495624
REFERENCE   2  (bases 1 to 6696)
  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: 
            "10.1038/s41564-020-00853-0"; journalName: "Nat Microbiol"; date: 
            "2021-05"; volume: "6"; issue: "5"; pages: "584-593"
COMMENT     Sequence Label: pXY349
FEATURES             Location/Qualifiers
     source          1..6696
                     /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             1478..2206
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDM
                     ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     terminator      2258..2352
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2422..2444)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(2909..3700,3701..3769))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3532..3551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3770..3874)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4425..5013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4914..4933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5167..5184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5257..5334
                     /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             5335..6417
                     /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(5354..5373)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6466..6471,6472..6489,6490..6496)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6504..6520
                     /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     6528..6544
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6528..6544
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6560..6577)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6560..6576)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6569..6591)
                     /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 atgcgtttat ctctccctcg cctgaaaatg ccgcgcctgc caggattcag
      241 tatcctggtc tggatctcca cggcgctaaa gggctgggtg atgggctcgc gggaaaaaga
      301 taccgacagc ctgatcatgt acgatcgcac cttactgtgg ctgaccttcg gcctcgcggc
      361 gattggcttt atcatggtga cctcggcgtc aatgcccata gggcaacgct taaccaacga
      421 tccgttcttc ttcgcgaagc gtgatggtgt ctatctgatt ttggcgttta ttctggcgat
      481 cattacgctg cgtctgccga tggagttctg gcaacgctac agtgccacga tgctgctcgg
      541 atctatcatc ctgctgatga tcgtcctggt agtgggtagc tcggttaaag gggcatcgcg
      601 ttggatcgat ctcggtttgc tgcgtatcca gcctgcggag ctgacaaaac tgtcgctgtt
      661 ttgctatatc gccaactatc tggtgcgtaa aggcgacgaa gtacgtaata acctgcgcgg
      721 cttcctgaaa ccgatgggcg tgattctggt gttggcagtg ttactgctgg cacagccaga
      781 ccttggtacg gtggtggtgt tgtttgtgac tacgctggcg atgttgttcc tggcgggagc
      841 gaaattgtgg cagttcattg ccattatcgg tatgggcatt tcagcggttg tgttgctgat
      901 actcgccgaa ccgtaccgta tccgccgtgt taccgcattc tggaacccgt gggaagatcc
      961 ctttggcagc ggctatcagt taacgcaatc gctgatggcg tttggtcgcg gcgaactttg
     1021 ggggcaaggt ttaggtaact cggtacaaaa actggagtat ctgccggaag cgcacactga
     1081 ctttattttc gccattatcg gcgaagaact ggggtatgtc ggtgtggtgc tggcactttt
     1141 aatggtattc ttcgtcgctt ttcgcgcgat gtcgattggc cgtaaagcat tagaaattga
     1201 ccaccgtttt tccggttttc tcgcctgttc tattggcatc tggtttagct tccaggcgct
     1261 ggttaacgta ggcgcggcgg cggggatgtt accgaccaaa ggtctgacat tgccgctgat
     1321 cagttacggt ggttcgagct tactgattat gtcgacagcc atcatgatgc tgttgcgtat
     1381 tgattatgaa acgcgtctgg agaaagcgca ggcgtttgta cgaggttcac gaggtggcgg
     1441 tggtagcccg gcaccggcac cgggcggtgg cggtagcgtg tctaagggcg aagagctgat
     1501 taaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
     1561 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
     1621 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     1681 cagcagaacc ttcatcaacc acacccaggg catccccgat ttctttaagc agtccttccc
     1741 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     1801 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     1861 cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccaacaccga
     1921 gatgctgtac cccgctgacg gcggcctgga aggcagaacc gacatggccc tgaagctcgt
     1981 gggcgggggc cacctgatct gcaacttcaa gaccacatac agatccaaga aacccgctaa
     2041 gaacctcaag atgcccggcg tctactatgt ggaccacaga ctggaaagaa tcaaggaggc
     2101 cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc
     2161 tagcaaactg gggcacaaac ttaatggcat ggacgagctg tacaagtaag cggccgctaa
     2221 gggtcgacct gcagccaagc ttaattagct gagcttggac tcctgttgat agatccagta
     2281 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     2341 attggtgaga atccaagcta ggggaattca tcgtgactga ctgacgatct gcctcgcgcg
     2401 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2461 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     2521 gtgtcggggc gcagccatga cccggtcacg tagcgatagc ggagtgtata attcttgaag
     2581 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2641 ttaggatccg ggatgtccgg cggtgctttt gccgttacgc accaccccgt cagtagctga
     2701 acaggaggga cagctgatag aaacagaagc cactggagca cctcaaaaac accatcatac
     2761 actaaatcag taagttggca gcatcacccg acgcactttg cgccgaataa atacctgtga
     2821 cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc gggaagccct
     2881 gggccaactt ttggcgaaaa tgagacgttt accaatgctt aatcagtgag gcacctatct
     2941 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3001 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3061 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3121 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3181 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3241 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3301 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3361 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3421 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3481 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3541 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3601 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3661 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3721 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3781 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3841 gtatttagaa aaataaacaa ataggggttc cgcgtttttt taaggcagtt attggtgccc
     3901 ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg gcagaaattc
     3961 gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc
     4021 tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat
     4081 gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacgt tatgatcatt
     4141 tattctgcct cccagagcct gataaaaacg gttaccggat cgagtcaggc aactatggat
     4201 gaacgaaata gacagatcgc ttagataggt gcctcactga ttaagcattg gtaactgtca
     4261 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4321 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4381 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4441 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4501 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4561 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4621 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4681 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac
     4741 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4801 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4861 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4921 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4981 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5041 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5101 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5161 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     5221 acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg ttgacaaacc
     5281 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     5341 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     5401 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     5461 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     5521 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     5581 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     5641 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     5701 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     5761 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     5821 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     5881 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     5941 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     6001 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     6061 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     6121 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     6181 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     6241 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     6301 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6361 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     6421 caacgcaact aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct
     6481 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     6541 tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     6601 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     6661 aagaggcccg caccgatcgc ccttcccaac agttgc
//
