LOCUS       sequence-502024-        7237 bp    DNA     circular SYN 05-MAR-2026
DEFINITION  mRFP-PopZ, sfGFP-PodJ249pse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7237)
  AUTHORS   Chou TW, Huang IJ, Hsu YC, Wu SH, Hong JC, Ko YH, Su TR, Jiang JR, 
            Huang HC
  TITLE     Construction and phenotypic classification of synthetic dual-pole 
            Escherichia coli cells.
  JOURNAL   Commun Biol. 2025 Jul 20;8(1):1078. doi: 10.1038/s42003-025-08495-w.
  PUBMED    40684034
REFERENCE   2  (bases 1 to 7237)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 5, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2025-07-20"; volume: "8(1):1078. doi"; pages: " 
            10.1038/s42003-025-08495-w"
COMMENT     Sequence Label: RP+ PodJ'
FEATURES             Location/Qualifiers
     source          1..7237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(29..56)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(72..143)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(162..179)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             complement(735..1409)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             1416..1427
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(1439..1473)
                     /label=BBa_J23116
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.16"
     terminator      complement(1509..1538)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             complement(1601..2683)
                     /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     2645..2664
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2818..2823,2824..2839,2840..2846)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2852..2871
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     RBS             2890..2901
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2910..2912,2913..3623)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2910..3623
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2940..2968)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3563..3583
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      5060..5131
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5147..5174
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      5211..5268
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(5292..5309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5463..6051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5543..5562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6234..6328)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6349..7008)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6923..6942
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     7170..7188
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7192..7235)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattccggc tctagagcag tatataaacg cagaaaggcc cacccgaagg tgagccagtg
       61 tgactctagt agagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
      121 gactgagcct ttcgttttat ttgatgcctg gctctagttt aacaacatcc tggacaacag
      181 gcgccgcgtc cccgagagat acgctgcact tcttcctcaa ccttggtctc gacgatgcgc
      241 ggcaggttct ggtccagcca ctccttgagc agcgggcgca gcagctcgcg tacgacgtct
      301 tccagcgtcc gaccgtcctt gggcatcagc agggccgagc tcaggctgcc gaaggcgctc
      361 gccgcggccg aagcggccga aacgccgacc agctgctcgg cgacttcgtc gcgatcaaac
      421 accggagccg ccggcggcgg cgtgtaggcc ggttccgact caggttccgg cggcgaatag
      481 acgtcgatgt cgccgacagt ctccagcggc ggcagctcgg gctcgggcgc gatcgggtcc
      541 gtcaattcca gaacctcgtc gtcgaacgac accggttcag gttcgggttc cggcggcggc
      601 gggggcgccg cttcggccgc aggctccgcc ggcgcgtcat cctccgagat gatgcgtcga
      661 atggaggcga ggatttcctc cattgtaggt tcttgagact gatcggacat ggatcccgat
      721 gagccagatg atccagcacc ggtggagtga cgaccttcag cacgttcgta ctgttcaacg
      781 atggtgtagt cttcgttgtg ggaggtgatg tccagtttga tgtcggtttt gtaagcaccc
      841 ggcagctgaa ccggtttttt agccatgtag gtggttttaa cttcagcgtc gtagtgacca
      901 ccgtctttca gtttcagacg cattttgatt tcacctttca gagcaccgtc ttccgggtac
      961 atacgttcgg tggaagcttc ccaacccatg gtttttttct gcataaccgg accgtcggac
     1021 gggaagttgg taccacgcag tttaactttg tagatgaact caccgtcttg cagggaggag
     1081 tcctgggtaa cggtaacaac accaccgtct tcgaagttca taacacgttc ccatttgaaa
     1141 ccttccggga aggacagttt caggtagtcc gggatgtcag ccgggtgttt aacgtaagct
     1201 ttggaaccgt actggaactg cggggacagg atgtcccaag cgaacggcag cggaccacct
     1261 ttggtaactt tcagtttagc ggtctgggta ccttcgtacg gacgaccttc accttcacct
     1321 tcgatttcga actcgtgacc gttaacggaa ccttccatac gaactttgaa acgcatgaac
     1381 tctttgataa cgtcttcgga ggaagccatc tagtatttct cctctttaat ctctagtagc
     1441 tagcatagtc cctaggactg agctagctgt caactctcgg actagaatat tcagcaattt
     1501 gcccgtgccg aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gctacgtaat
     1561 gtcggccaat tcgcgctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1621 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1681 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gactggcaac agctgattgc
     1741 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     1801 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagcta tcttcggtat
     1861 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     1921 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     1981 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     2041 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     2101 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     2161 ccagatgctc cacgcccagt cgcgtaccgt cctcatggga gaaaataata ctgttgatgg
     2221 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     2281 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     2341 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     2401 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     2461 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     2521 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     2581 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     2641 caccggcata ctctgcgaca tcgtataacg ttactggttt catattcacc accctgaatt
     2701 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggcgc
     2761 gccgcttcgt caggccacat agctttcttg ttctgatcgg aacgatcgtt ggctgtgttg
     2821 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cgctcacaat tctatggact
     2881 atactagaga aagaggagaa atactagaga tgagcaaagg agaagaactt ttcactggag
     2941 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtccgtg
     3001 gagagggtga aggtgatgct acaaacggaa aactcaccct taaatttatt tgcactactg
     3061 gaaaactacc tgttccgtgg ccaacacttg tcactactct gacctatggt gttcaatgct
     3121 tttcccgtta tccggatcac atgaaacggc atgacttttt caagagtgcc atgcccgaag
     3181 gttatgtaca ggaacgcact atatctttca aagatgacgg gacctacaag acgcgtgctg
     3241 aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaagggt attgatttta
     3301 aagaagatgg aaacattctt ggacacaaac tcgagtacaa ctttaactca cacaatgtat
     3361 acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt cgccacaacg
     3421 ttgaagatgg ttccgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     3481 gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgtcctt tcgaaagatc
     3541 ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct gggattacac
     3601 atggcatgga tgagctctac aaatgttttc acacaggaaa ggcctcgatg actgctgctt
     3661 cgccatggag cgttaagggg attgacccca aggcacggga ggtcgcgaaa gacctcgcgc
     3721 gtcgctccgg catgactttg ggtgaatggc tgaaccgaat gatcatcgaa ggcgatggtc
     3781 agaccgctga tccgcgtctc gcgggggacg atgtccccaa ccgcgcctat ctcgagattg
     3841 tcaaagatga tgcgccgccg cgcatcgaga tcgccgagca tccggcagat gaggttggtc
     3901 gcgtcgcgct ggcgctggat cgcctgaccc aacgcattga agccgccgag gggcgcaacg
     3961 ccgccgccat caccggcatc gatcactccg tccgtgacgc cctgacgcgt ctgggcgcat
     4021 ccgaacgtga gcagatcgcc gtcgccgccc ggttcgaggg cgctgtcgac gaactgaaga
     4081 ccgagcaggc tcgggccacc gaacgtctgc gccggatcga gagcgaggcg gcgggtccgc
     4141 gctcggccga ggcgttgcgc gcgcttgaag gcgccttggg caaggtcgcc ggacaccttt
     4201 acgaaggcga ggcgcggacg cgcgaggcga tcgcgaccct ggaagccaag ctgaaccagc
     4261 agagctcggg cgatcccagc gcgctggtcg aggctgtggt cgcgcgactg ggcgaacgcc
     4321 tcgaagccgc cgaaacccgc accagcgacg cgctgcgcga actgggcgcg tcgttccagg
     4381 ccctggatca gcgccaggaa ctggtcgacc gcatccgcca gagcgaagag cgcacgctcc
     4441 gcatgctgga agaagcgcgc gagaagatcg acagccgtct gtccgaggcg cagcggaagc
     4501 tggaagctgc gccgccgtcg ccgccgcccg cgcaggctcc ggcgccggtt gcgactgctc
     4561 agcgtccggt tccgccggcg gcttcgccct tcgaggacaa ctatttcagc caggccgcct
     4621 cgttcagcac gtctgaggat gaggctgacg cgtttgatgc gccgcctgcg ccggcccgta
     4681 gcttcgaggt ggcggagttc ccggctgcgg agccggaaga accggccttc gcacacgatg
     4741 actacgcgat cgccgatggc ttcgagccgg aaagcccgcg ctatgaagtc gagcctgagg
     4801 tctccgactt cgcgcccgcc gagccgtcgc gtccgatgtc gacccgcgat atcatcgagc
     4861 aggcccgcgc cgccgcgcgc gccgccgccg cctcggaagg caagggcggc aaggccaagt
     4921 cggcgaagaa ggaaaaggct tccaaggcca gcggctcgct gttcagcggc tttggcggtt
     4981 tctcgaccaa gaagtcgaag gcgcgcttgg gcgcgaccgt gacgacggcc taataactag
     5041 ggcccatacc cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     5101 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     5161 tgggcctttc tgcgtttata cgacgtacgg actagtagcg gccgctgcag tccggcaaaa
     5221 aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt
     5281 atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5341 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5401 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5461 tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5521 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5581 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5641 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5701 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5761 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5821 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5881 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5941 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6001 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6061 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gctcgaggct tggattctca
     6241 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     6301 gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc
     6361 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     6421 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     6481 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     6541 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     6601 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     6661 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     6721 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     6781 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     6841 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     6901 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     6961 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     7021 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     7081 gtgaaagttg gaacctctta cgtgcccgat caactcgagt gccacctgac gtctaagaaa
     7141 ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat
     7201 aaaaaaaatc cttagctttc gctaaggatg atttctg
//
