LOCUS       sequence-266114-        6221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6221)
  AUTHORS   Brodel AK, Rodrigues R, Jaramillo A, Isalan M
  TITLE     Accelerated evolution of a minimal 63-amino acid dual transcription 
            factor.
  JOURNAL   Sci Adv. 2020 Jun 10;6(24):eaba2728. doi: 10.1126/sciadv.aba2728. 
            eCollection 2020 Jun.
  PUBMED    32577520
REFERENCE   2  (bases 1 to 6221)
  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: "Sci Adv."; 
            date: "2020-06-10"; volume: "6(24):eaba2728. doi"; pages: " 
            10.1126/sciadv.aba2728. eCollection 2020 Jun"
COMMENT     Sequence Label: pJPC12-Delta-PS-mCherry-PCS/PM,CS-GFP
FEATURES             Location/Qualifiers
     source          1..6221
                     /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             1520..1531
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1538..2251
                     /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(2289..2306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      3451..3673
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3721..4671
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(5708..6221,1..146))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
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 ccggcggtga taaaatagtc aacaccggcg gtgatagata tttataaata
     1501 gtggtgatag atttaacgta aagaggagaa atactagatg cgtaaaggcg aagaactgtt
     1561 taccggtgtg gttccgattc tggtggaact ggacggcgat gttaatggtc ataaattcag
     1621 tgttcgcggc gaaggtgaag gcgatgcgac gaacggcaaa ctgaccctga aatttatctg
     1681 caccacgggt aaactgccgg tcccgtggcc gacgctggtg accacgctga cctatggcgt
     1741 tcaatgtttt gcgcgttacc cggatcacat gaaacagcac gactttttca aatcggccat
     1801 gccggaaggc tatgtgcagg aacgtacgat tagctttaaa gacgatggta cgtataaaac
     1861 ccgcgcggaa gtgaaattcg aaggcgatac cctggttaac cgtatcgaac tgaaaggtat
     1921 cgatttcaaa gaagacggca atattctggg tcataaactg gaatataact tcaattccca
     1981 caacgtgtac atcaccgcgg ataaacagaa aaacggcatt aaagccaatt tcaaaatccg
     2041 ccataatgtg gaagatggta gcgttcagct ggccgaccac tatcagcaaa acacgccgat
     2101 tggtgatggc ccggtcctgc tgccggacaa tcactacctg agtacccagt ccgtgctgtc
     2161 aaaagatccg aacgaaaaac gtgaccacat ggtcctgctg gaatttgtga cggctgcggg
     2221 tatcacccac ggcatggacg aactgtataa ataacgcaaa aaaccccgct tcggcggggt
     2281 tttttcgcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2401 atgtgagatc tctacgggtc ggatttgaag tcgtcttggt aggaggcagc ctgaatggcg
     2461 aatgccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     2521 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacagggtgc
     2581 cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg
     2641 gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc
     2701 ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc
     2761 tgggatacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc
     2821 ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag
     2881 atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga
     2941 tcattggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt
     3001 tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg
     3061 catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac
     3121 cactccgcag acccgccata aaacgccctg agaagcccgt gacgggcttt tcttgtatta
     3181 tgggtagttt ccttgcatga atccataaaa ggcgcctgta gtgccattta cccccattca
     3241 ctgccagagc cgtgagcgca gcgaactgaa tgtcacgaaa aagacagcga ctcaggtgcc
     3301 tgatggtcgg agacaaaagg aatattcagc gatttgcccg agcttgcgag ggtgctactt
     3361 aagcctttag ggttttaagg tctgttttgt agaggagcaa acagcgtttg cgacatcctt
     3421 ttgtaatact gcggaactga ctaaagtagt gagttataca cagggctggg atctattctt
     3481 tttatctttt tttattcttt ctttattcta taaattataa ccacttgaat ataaacaaaa
     3541 aaaacacaca aaggtctagc ggaatttaca gagggtctag cagaatttac aagttttcca
     3601 gcaaaggtct agcagaattt acagataccc acaactcaaa ggaaaaggac tagtaattat
     3661 cattgactag cccatctcaa ttggtatagt gattaaaatc acctagacca attgagatgt
     3721 atgtctgaat tagttgtttt caaagcaaat gaactagcga ttagtcgcta tgacttaacg
     3781 gagcatgaaa ccaagctaat tttatgctgt gtggcactac tcaaccccac gattgaaaac
     3841 cctacaagga aagaacggac ggtatcgttc acttataacc aatacgctca gatgatgaac
     3901 atcagtaggg aaaatgctta tggtgtatta gctaaagcaa ccagagagct gatgacgaga
     3961 actgtggaaa tcaggaatcc tttggttaaa ggctttaaga tcttccagtg gacaaactat
     4021 gccaagttct caagcgaaaa attagaatta gtttttagtg aagagatatt gccttatctt
     4081 ttccagttaa aaaaattcat aaaatataat ctggaacatg ttaagtcttt tgaaaacaaa
     4141 tactctatga ggatttatga gtggttatta aaagaactaa cacaaaagaa aactcacaag
     4201 gcaaatatag agattagcct tgatgaattt aagttcatgt taatgcttga aaataactac
     4261 catgagttta aaaggcttaa ccaatgggtt ttgaaaccaa taagtaaaga tttaaacact
     4321 tacagcaata tgaaattggt ggttgataag cgaggccgcc cgactgatac gttgattttc
     4381 caagttgaac tagatagaca aatggatctc gtaaccgaac ttgagaacaa ccagataaaa
     4441 atgaatggtg acaaaatacc aacaaccatt acatcagatt cctacctaca taacggacta
     4501 agaaaaacac tacacgatgc tttaacagca aaaattcagc tcaccagttt tgaggcaaaa
     4561 tttttgagtg acatgcaaag taagtatgat ctcaatggtt cgttctcatg gctcacgcaa
     4621 aaacaacgaa ccacactaga gaacatactg gctaaatacg gaaggatctg aggttcttat
     4681 ggctcttgta tctatcagtg aagcatcaag actaacaaac aaaagtagaa caactgttca
     4741 ccgttacata tcaaagggaa aactgtccat atgcacagat gaaaacggtg taaaaaagat
     4801 agatacatca gagcttttac gagtttttgg tgcattcaaa gctgttcacc atgaacagat
     4861 cgacaatgta acagatgaac agcatgtaac acctaataga acaggtgaaa ccagtaaaac
     4921 aaagcaacta gaacatgaaa ttgaacacct gagacaactt gttacagctc aacagtcaca
     4981 catagatagc ctgaaacagg cgatgctgct tatcgaatca aagctgccga caacacggga
     5041 gccagtgacg cctcccgtgg ggaaaaaatc atggcaattc tggaagaaat agcgctttca
     5101 gccggcaaac cggctgaagc cggatctgcg attctgataa caaactagca acaccagaac
     5161 agcccgtttg cgggcagcaa aacccgtact tttggacgtt ccggcggttt tttgtggcga
     5221 gtggtgttcg ggcggtgcgc gcaagatcca ttatgttaaa cgggcgagtt tacatctcaa
     5281 aaccgcccgc ttaacaccat cagaaatcct cagcgcgatt ttaagcacca accccccccg
     5341 taacacccaa atccatactg aaagtggctt tgttgaataa atcagatttc gggtaagtct
     5401 cccccgtagc gggttgtgtt ttcaggcaat acgcacgctt tcaggcatac ctgctttcgt
     5461 cattttgttc agcgctcgta ccagggccat agcctccgca acctgaccat cgtagtcacg
     5521 cagcgtcagt gaacccccga acagagatct gacgctcagt ggaacgaaaa ctcacgttaa
     5581 gggattttgg tcatgagaca ataaccctga taaatgcttc aataatcgca gaccaaaacg
     5641 atctcaagaa gatcatctta ttaatcagat aaaatatttc taggcaccaa taactgcctt
     5701 aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     5761 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     5821 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     5881 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     5941 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     6001 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     6061 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     6121 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     6181 gaatgtgaat aaaggccgga taaaacttgt gcttattttt c
//
