LOCUS       sequence-207939-        5431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric reporter [PT7(-3G)-RBST7g10, high copy number].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5431)
  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 5431)
  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: p1eng3VT-lacI
FEATURES             Location/Qualifiers
     source          1..5431
                     /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"
     RBS             875..897
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(909..911,912..914,915..1628)
                     /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      1648..1719
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1735..1762
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1769..1803
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1812..1823
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1839..2918
                     /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(1858..1877)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2919..2951
                     /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      2999..3070
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3086..3113
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3120..3140
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(join(3166..3954,3955..4023))
                     /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     3786..3805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4024..4128)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4196..4214
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4218..4261)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4264..4285
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      complement(4292..4319)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4335..4406)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(4426..5139,5140..5142,5143..5145))
                     /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             5152..5163
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(5172..5206)
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     misc_feature    5207..5227
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5228..5285
                     /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(5309..5326)
                     /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 actagaggcg aaattaatac gactcactgt
      841 agggagacca caacggtttc cctctagaaa taattttgtt taactttaag aaggagatat
      901 actactagat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      961 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1021 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     1081 ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc
     1141 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1201 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1261 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1321 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     1381 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     1441 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1501 acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1561 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1621 acaagtaata atactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1681 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     1741 ccttcgggtg ggcctttctg cgtttatatt tacagctagc tcagtcctag gttttatgct
     1801 agctactaga gaaagaggag aaatactaga tggtgaatgt gaaaccagta acgttatacg
     1861 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     1921 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     1981 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     2041 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     2101 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     2161 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     2221 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     2281 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     2341 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     2401 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     2461 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     2521 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     2581 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     2641 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
     2701 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     2761 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     2821 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2881 agctggcacg acaggtttcc cgactggaaa gcgggcaggc tgcaaacgac gaaaactacg
     2941 ctttagtagc ttaataactc tgatagtgct agtgtagatc tctactagag ccaggcatca
     3001 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3061 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat
     3121 actagtagcg gccgctgcag tgatctacac tagcactatc agtgaccaat gcttaatcag
     3181 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3241 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3301 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3361 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3421 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3481 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3541 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3601 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3661 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3721 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3781 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3841 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3901 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3961 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4021 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4081 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4141 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     4201 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
     4261 ctggaattcg cggccgcttc tagagtataa acgcagaaag gcccacccga aggtgagcca
     4321 gtgtgactct agtagagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     4381 ttcgactgag cctttcgttt tatttgatgc ctggctctag tattatcact tgtacagctc
     4441 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
     4501 cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag
     4561 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc
     4621 gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct tctgcttgtc
     4681 ggcggtgata tagacgttgt cgctaaagta gttgtactcc agcttgtgcc ccaggatgtt
     4741 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     4801 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     4861 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     4921 gtagcgggcg aagcactgca cgccccagga cagggtggtc acgagggtgg gccagggcac
     4981 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
     5041 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     5101 gatgggcacc accccggtga acagctcctc gcccttgctc accatctagt atttctcctc
     5161 tttctctagt agctagcata atacctagga ctgagctagc tgtaaatact agtagcggcc
     5221 gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct ttaaaaccga
     5281 aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5341 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5401 gggataacgc aggaaagaac atgtgagcaa a
//
