LOCUS       sequence-152669-        6238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid (mCherry and GFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6238)
  AUTHORS   Brodel AK, Jaramillo A, Isalan M
  TITLE     Engineering orthogonal dual transcription factors for multi-input 
            synthetic promoters.
  JOURNAL   Nat Commun. 2016 Dec 16;7:13858. doi: 10.1038/ncomms13858.
  PUBMED    27982027
REFERENCE   2  (bases 1 to 6238)
  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: "2016-12-16"; volume: "7:13858. doi"; pages: " 
            10.1038/ncomms13858"
COMMENT     Sequence Label: pJPC12-Delta-M13-mCherry-PR/PRM-GFP
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..80
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     471..493
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     485..502
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     486..502
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     512..531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        512..530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     555..571
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     556..572
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(607..693)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(709..1419)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(987..1006)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1256..1274
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             1426..1437
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             1531..1542
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1549..2262
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2300..2317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      3462..3684
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3732..4682
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
       61 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
      121 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
      181 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttaca
      241 aaaggatcta ggtgaagatc ctttttgata atctcatgag cggatacata tttgaatgta
      301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccaccttcag
      361 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      421 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      481 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
      541 taccgggccc cccctcgagg tcgacggtat cgataagctt gatatcgaat tcgtctaggg
      601 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
      661 ccagtctttc gactgagcct ttcgttttat ttgatgcctc tactcgagtt acttgtacag
      721 ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt
      781 gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag
      841 ctgcacgggc ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc
      901 cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg
      961 ctcggaggag gcctcccagc ccatagtctt cttctgcatt acggggccgt cggaggggaa
     1021 gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg
     1081 ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc
     1141 ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga
     1201 gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt
     1261 caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat
     1321 ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt
     1381 gatgatagcc atgttatcct cctcgccctt gctcaccatg aattctttct cctctttgca
     1441 accattatca ccgccagagg taaaatagtc aacacgcacg gtgttagata tttataaata
     1501 gtggtgatag atttaacgtt actagagatt aaagaggaga aatactagat gcgtaaaggc
     1561 gaagaactgt ttaccggtgt ggttccgatt ctggtggaac tggacggcga tgttaatggt
     1621 cataaattca gtgttcgcgg cgaaggtgaa ggcgatgcga cgaacggcaa actgaccctg
     1681 aaatttatct gcaccacggg taaactgccg gtcccgtggc cgacgctggt gaccacgctg
     1741 acctatggcg ttcaatgttt tgcgcgttac ccggatcaca tgaaacagca cgactttttc
     1801 aaatcggcca tgccggaagg ctatgtgcag gaacgtacga ttagctttaa agacgatggt
     1861 acgtataaaa cccgcgcgga agtgaaattc gaaggcgata ccctggttaa ccgtatcgaa
     1921 ctgaaaggta tcgatttcaa agaagacggc aatattctgg gtcataaact ggaatataac
     1981 ttcaattccc acaacgtgta catcaccgcg gataaacaga aaaacggcat taaagccaat
     2041 ttcaaaatcc gccataatgt ggaagatggt agcgttcagc tggccgacca ctatcagcaa
     2101 aacacgccga ttggtgatgg cccggtcctg ctgccggaca atcactacct gagtacccag
     2161 tccgtgctgt caaaagatcc gaacgaaaaa cgtgaccaca tggtcctgct ggaatttgtg
     2221 acggctgcgg gtatcaccca cggcatggac gaactgtata aataacgcaa aaaaccccgc
     2281 ttcggcgggg ttttttcgcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2341 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2401 caggaaagaa catgtgagat ctctacgggt cggatttgaa gtcgtcttgg taggaggcag
     2461 cctgaatggc gaatgccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     2521 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     2581 ggacagggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     2641 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     2701 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     2761 ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga tggccttccc
     2821 cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc
     2881 caggcaggta gatgacgacc atcagggaca gcttcaagga tcgctcgcgg ctcttaccag
     2941 cctaacttcg atcattggac cgctgatcgt cacggcgatt tatgccgcct cggcgagcac
     3001 atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt
     3061 gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccggcg gcacctcgct
     3121 aacggattca ccactccgca gacccgccat aaaacgccct gagaagcccg tgacgggctt
     3181 ttcttgtatt atgggtagtt tccttgcatg aatccataaa aggcgcctgt agtgccattt
     3241 acccccattc actgccagag ccgtgagcgc agcgaactga atgtcacgaa aaagacagcg
     3301 actcaggtgc ctgatggtcg gagacaaaag gaatattcag cgatttgccc gagcttgcga
     3361 gggtgctact taagccttta gggttttaag gtctgttttg tagaggagca aacagcgttt
     3421 gcgacatcct tttgtaatac tgcggaactg actaaagtag tgagttatac acagggctgg
     3481 gatctattct ttttatcttt ttttattctt tctttattct ataaattata accacttgaa
     3541 tataaacaaa aaaaacacac aaaggtctag cggaatttac agagggtcta gcagaattta
     3601 caagttttcc agcaaaggtc tagcagaatt tacagatacc cacaactcaa aggaaaagga
     3661 ctagtaatta tcattgacta gcccatctca attggtatag tgattaaaat cacctagacc
     3721 aattgagatg tatgtctgaa ttagttgttt tcaaagcaaa tgaactagcg attagtcgct
     3781 atgacttaac ggagcatgaa accaagctaa ttttatgctg tgtggcacta ctcaacccca
     3841 cgattgaaaa ccctacaagg aaagaacgga cggtatcgtt cacttataac caatacgctc
     3901 agatgatgaa catcagtagg gaaaatgctt atggtgtatt agctaaagca accagagagc
     3961 tgatgacgag aactgtggaa atcaggaatc ctttggttaa aggctttaag atcttccagt
     4021 ggacaaacta tgccaagttc tcaagcgaaa aattagaatt agtttttagt gaagagatat
     4081 tgccttatct tttccagtta aaaaaattca taaaatataa tctggaacat gttaagtctt
     4141 ttgaaaacaa atactctatg aggatttatg agtggttatt aaaagaacta acacaaaaga
     4201 aaactcacaa ggcaaatata gagattagcc ttgatgaatt taagttcatg ttaatgcttg
     4261 aaaataacta ccatgagttt aaaaggctta accaatgggt tttgaaacca ataagtaaag
     4321 atttaaacac ttacagcaat atgaaattgg tggttgataa gcgaggccgc ccgactgata
     4381 cgttgatttt ccaagttgaa ctagatagac aaatggatct cgtaaccgaa cttgagaaca
     4441 accagataaa aatgaatggt gacaaaatac caacaaccat tacatcagat tcctacctac
     4501 ataacggact aagaaaaaca ctacacgatg ctttaacagc aaaaattcag ctcaccagtt
     4561 ttgaggcaaa atttttgagt gacatgcaaa gtaagtatga tctcaatggt tcgttctcat
     4621 ggctcacgca aaaacaacga accacactag agaacatact ggctaaatac ggaaggatct
     4681 gaggttctta tggctcttgt atctatcagt gaagcatcaa gactaacaaa caaaagtaga
     4741 acaactgttc accgttacat atcaaaggga aaactgtcca tatgcacaga tgaaaacggt
     4801 gtaaaaaaga tagatacatc agagctttta cgagtttttg gtgcattcaa agctgttcac
     4861 catgaacaga tcgacaatgt aacagatgaa cagcatgtaa cacctaataa gaacaggtga
     4921 aaccagtaaa acaaagcaac tagaacatga aattgaacac ctgagacaac ttgttacagc
     4981 tcaacagtca cacatagaca gcctgaaaca ggcgatgctg cttatcgaat caaagctgcc
     5041 gacaacacgg gagccagtga cgcctcccgt ggggaaaaaa tcatggcaat tctggaagaa
     5101 atagcgcttt cagccggcaa accggctgaa gccggatctg cgattctgat aacaaactag
     5161 caacaccaga acagcccgtt tgcgggcagc aaaacccgta cttttggacg ttccggcggt
     5221 tttttgtggc gagtggtgtt cgggcggtgc gcgcaagatc cattatgtta aacgggcgag
     5281 tttacatctc aaaaccgccc gcttaacacc atcagaaatc ctcagcgcga ttttaagcac
     5341 caaccccccc ccgtaacacc caaatccata ctgaaagtgg ctttgttgaa taaatcagat
     5401 ttcgggtaag tctcccccgt agcgggttgt gttttcaggc aatacgcacg ctttcaggca
     5461 tacctgcttt cgtcattttg ttcagcgctc gtaccagggc catagcctcc gcaacctgac
     5521 catcgtagtc acgcagcgtc agtgaacccc cgaacagaga tctgacgctc agtggaacga
     5581 aaactcacgt taagggattt tggtcatgag acaataaccc tgataaatgc ttcaataatc
     5641 gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat ttctaggcac
     5701 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     5761 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     5821 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     5881 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     5941 gctgaaacga aaaacatatt ctcaataaac ccttttaggg aaataaggcc aggttttcac
     6001 cgtaacacgc cacatcttgg gaaatatttg tgtagaaact gccggaaatc gtcgtggtat
     6061 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     6121 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     6181 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcc
//
