LOCUS       sequence-207686-        6644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric reporter [PT7-RBSB0034, low copy number].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6644)
  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 6644)
  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: p4wtVT-lacI
FEATURES             Location/Qualifiers
     source          1..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             19..969
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTDYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     primer_bind     1179..1198
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1179..1197
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1233..1244
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1251..1253,1254..1256,1257..1970)
                     /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      1990..2061
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2077..2104
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2111..2145
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             2154..2165
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2181..3260
                     /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(2200..2219)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3261..3293
                     /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      3341..3412
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3428..3455
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3462..3482
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             join(3586..3654,3655..4446)
                     /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"
     terminator      complement(4576..4605)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(4680..4707)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4723..4794)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(4814..5527,5528..5530,5531..5533))
                     /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             5540..5551
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(5560..5594)
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     misc_feature    5595..5615
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5651..5682
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      5807..5901
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6393..6615
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ctagaccaat tgagatgtat gtctgaatta gttgttttca aagcaaatga actagcgatt
       61 agtcgctatg acttaacgga gcatgaaacc aagctaattt tatgctgtgt ggcactactc
      121 aaccccacga ttgaaaaccc tacaaggaaa gaacggacgg tatcgttcac ttataaccaa
      181 tacgctcaga tgatgaacat cagtagggaa aatgcttatg gtgtattagc taaagcaacc
      241 agagagctga tgacgagaac tgtggaaatc aggaatcctt tggttaaagg ctttgagatt
      301 ttccagtgga cagactatgc caagttctca agcgaaaaat tagaattagt ttttagtgaa
      361 gagatattgc cttatctttt ccagttaaaa aaattcataa aatataatct ggaacatgtt
      421 aagtcttttg aaaacaaata ctctatgagg atttatgagt ggttattaaa agaactaaca
      481 caaaagaaaa ctcacaaggc aaatatagag attagccttg atgaatttaa gttcatgtta
      541 atgcttgaaa ataactacca tgagtttaaa aggcttaacc aatgggtttt gaaaccaata
      601 agtaaagatt taaacactta cagcaatatg aaattggtgg ttgataagcg aggccgcccg
      661 actgatacgt tgattttcca agttgaacta gatagacaaa tggatctcgt aaccgaactt
      721 gagaacaacc agataaaaat gaatggtgac aaaataccaa caaccattac atcagattcc
      781 tacctacata acggactaag aaaaacacta cacgatgctt taactgcaaa aattcagctc
      841 accagttttg aggcaaaatt tttgagtgac atgcaaagta agtatgatct caatggttcg
      901 ttctcatggc tcacgcaaaa acaacgaacc acactagaga acatactggc taaatacgga
      961 aggatctgag gttcttatgg ctcttgtatc tatcagtgaa gcatcaagac taacaaacaa
     1021 aagtagaaca actgttcacc gttacatatc aaagggaaaa ctgtccatat gcacagatga
     1081 aaacggtgta aaaaagatag atacatcaga gcttttacga gtttttggtg tattcaaagc
     1141 tgttcaccat gaacagatcg acaatgtaac tactagagta atacgactca ctatagggaa
     1201 tacaagctac ttgttctttt tgcatactag agaaagagga gaaatactag atggtgagca
     1261 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1321 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1381 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1441 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1501 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1561 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1621 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1681 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1741 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1801 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1861 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1921 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa taatactaga
     1981 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     2041 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     2101 tgcgtttata tttacagcta gctcagtcct aggttttatg ctagctacta gagaaagagg
     2161 agaaatacta gatggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     2221 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     2281 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     2341 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     2401 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     2461 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     2521 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     2581 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     2641 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     2701 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     2761 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     2821 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     2881 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     2941 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     3001 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     3061 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     3121 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     3181 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3241 cccgactgga aagcgggcag gctgcaaacg acgaaaacta cgctttagta gcttaataac
     3301 tctgatagtg ctagtgtaga tctctactag agccaggcat caaataaaac gaaaggctca
     3361 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     3421 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atactagtag cggccgctgc
     3481 agtgatctac actagcacta tcagtgctaa atacattcaa atatctatcc gctcatgaga
     3541 caataaccct gataaatgct tcaataatat tgaaaaagga agaatatgag tattcaacat
     3601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3661 gaaacgctgg tgaaagtaaa agatgccgaa gatcagttgg gtgcacgtgt gggttacatc
     3721 gaactggacc tcaacagcgg taagattctt gagagttttc gccccgaaga acgtttccca
     3781 atgatgagca cttttaaagt tctgctctgt ggcgcggtat tatcccgtat tgacgccggg
     3841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3901 gtcacagaaa agcatcttac ggacggcatg acagtacgcg aattatgcag cgctgccata
     3961 accatgagtg ataacacggc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4021 cttaccgctt ttttgcacaa catgggtgat catgtaactc gccttgatcg ttgggaaccg
     4081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agctatggca
     4141 acaacgttgc gcaaactctt aactggcgaa cttcttactc tcgcttcccg gcaacaatta
     4201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtcccgcgg tattattgca
     4321 gccctggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagccag
     4381 gcaactatgg acgaacgtaa tcgccagatc gctgagatag gtgcctccct gattaagcat
     4441 tggtaataat actagctccg gcaaaaaaac gggcaaggtg tcaccaccct gccctttttc
     4501 tttaaaaccg aaaagattac ttcgcgtttg ccacctgacg tctaagaaaa ggaatattca
     4561 gcaatttgcc cgtgccgaag aaaggcccac ccgtgaaggt gagccagtga gttgattgct
     4621 acgtaattag ttagttagcc cttagtgact cgaattcgct gccgcttcta gagtataaac
     4681 gcagaaaggc ccacccgaag gtgagccagt gtgactctag tagagagcgt tcaccgacaa
     4741 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     4801 ggctctagta ttatcacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     4861 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact
     4921 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     4981 gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt
     5041 tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtcg ctaaagtagt
     5101 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     5161 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     5221 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     5281 agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg ccccaggaca
     5341 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     5401 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     5461 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     5521 ccttgctcac catctagtat ttctcctctt tctctagtag ctagcataat acctaggact
     5581 gagctagctg taaatactag tagcggccgc tgcaggagtc actaagggtt agttagttag
     5641 attagcagaa agtcaaaagc ctccgaccgg aggcttttga ctaaaacttc ccttggggtt
     5701 atcattgggg ctcactcaaa ggcggtaatc agataaaaaa aatccttagc tttcgctaag
     5761 gatgatttct gctagagctg tcagaccaag tttacgagct cgcttggact cctgttgata
     5821 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     5881 gcgcttttta ttggtgagaa tccaagcact agggacagta agacgggtaa gcctgttgat
     5941 gataccgctg ccttactggg tgcattagcc agtctgaatg acctgtcacg ggataatccg
     6001 aagtggccag actggaaaat cagagggcag gaactgctga acagcaaaaa gtcagatagc
     6061 accacatagc agacccgcca taaaacgccc tgagaagccc gtgacgggct tttcttgtat
     6121 tatgggtagt ttccttgcat gaatccataa aaggcgcctg tagtgccatt tacccccatt
     6181 cactgccaga gccgtgagcg cagcgaactg aatgtcacga aaaagacagc gactcaggtg
     6241 cctgatggtc ggagacaaaa ggaatattca gcgatttgcc cgagcttgcg agggtgctac
     6301 ttaagccttt agggttttaa ggtctgtttt gtagaggagc aaacagcgtt tgcgacatcc
     6361 ttttgtaata ctgcggaact gactaaagta gtgagttata cacagggctg ggatctattc
     6421 tttttatctt tttttattct ttctttattc tataaattat aaccacttga atataaacaa
     6481 aaaaaacaca caaaggtcta gcggaattta cagagggtct agcagaattt acaagttttc
     6541 cagcaaaggt ctagcagaat ttacagatac ccacaactca aaggaagagg acatgtaatt
     6601 atcattgact agcccatctc aattggtata gtgattaaaa tcac
//
