LOCUS       sequence-203731-        5412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric reporter [PT7-RBSB0034, high copy number].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  AUTHORS   Boehm CR, Grant PK, Haseloff J
  TITLE     Programmed hierarchical patterning of bacterial populations.
  JOURNAL   Nat Commun. 2018 Feb 22;9(1):776. doi: 10.1038/s41467-018-03069-3.
  PUBMED    29472537
REFERENCE   2  (bases 1 to 5412)
  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: "Nat 
            Commun."; date: "2018-02-22"; volume: "9(1):776. doi"; pages: " 
            10.1038/s41467-018-03069-3"
COMMENT     Sequence Label: p1wtVT-lacI
FEATURES             Location/Qualifiers
     source          1..5412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(49..637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(129..148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     818..837
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        818..836
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             872..883
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(890..892,893..895,896..1609)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      1629..1700
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1716..1743
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1750..1784
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1793..1804
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1820..2899
                     /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(1839..1858)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2900..2932
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      2980..3051
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3067..3094
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3101..3121
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(join(3147..3935,3936..4004))
                     /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     3767..3786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4005..4109)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4177..4195
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4199..4242)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4245..4266
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      complement(4273..4300)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4316..4387)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(4407..5120,5121..5123,5124..5126))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     RBS             5133..5144
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(5153..5187)
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     misc_feature    5188..5208
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5209..5266
                     /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(5290..5307)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct
       61 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      121 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      181 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      241 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      301 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      361 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      421 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      481 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      541 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      601 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      661 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      721 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      781 tatatatgag taaacttggt ctgacagttt actagagtaa tacgactcac tatagggaat
      841 acaagctact tgttcttttt gcatactaga gaaagaggag aaatactaga tggtgagcaa
      901 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      961 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1021 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1081 cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1141 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1201 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1261 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1321 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1381 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     1441 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     1501 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1561 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat aatactagag
     1621 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1681 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
     1741 gcgtttatat ttacagctag ctcagtccta ggtattatgc tagctactag agaaagagga
     1801 gaaatactag atggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
     1861 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
     1921 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
     1981 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
     2041 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
     2101 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
     2161 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
     2221 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
     2281 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
     2341 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
     2401 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
     2461 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
     2521 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
     2581 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
     2641 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
     2701 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     2761 gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac
     2821 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     2881 ccgactggaa agcgggcagg ctgcaaacga cgaaaactac gctttagtag cttaataact
     2941 ctgatagtgc tagtgtagat ctctactaga gccaggcatc aaataaaacg aaaggctcag
     3001 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     3061 cacactggct caccttcggg tgggcctttc tgcgtttata tactagtagc ggccgctgca
     3121 gtgatctaca ctagcactat cagtgaccaa tgcttaatca gtgaggcacc tatctcagcg
     3181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3481 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3721 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3781 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3841 aggatcttac cgctgttgag atccagttcg atataaccca ctcgtgcacc caactgatct
     3901 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3961 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4021 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4081 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4141 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat
     4201 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt
     4261 ctagagtata aacgcagaaa ggcccacccg aaggtgagcc agtgtgactc tagtagagag
     4321 cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     4381 ttatttgatg cctggctcta gtattattac ttgtacagct cgtccatgcc gagagtgatc
     4441 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     4501 ctcagcttgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     4561 atgggggtgt tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg
     4621 cggatcttga agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg
     4681 tcgctaaagt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     4741 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     4801 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     4861 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     4921 acgccccagg acagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     4981 cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     5041 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     5101 aacagctcct cgcccttgct caccatctag tatttctcct ctttctctag tagctagcat
     5161 aatacctagg actgagctag ctgtaaatac tagtagcggc cgctgcagtc cggcaaaaaa
     5221 gggcaaggtg tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgttat
     5281 gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5401 catgtgagca aa
//
