LOCUS       sequence-103751-        4304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A reporter plasmid containing the attB and attP sites flanking a 
            green fluorescent protein reporter gene (gfp). attP upstream of attB
            in this plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4304)
  AUTHORS   Yang L, Nielsen AA, Fernandez-Rodriguez J, McClune CJ, Laub MT, Lu 
            TK, Voigt CA
  TITLE     Permanent genetic memory with >1-byte capacity.
  JOURNAL   Nat Methods. 2014 Dec;11(12):1261-6. doi: 10.1038/nmeth.3147. Epub 
            2014 Oct 26.
  PUBMED    25344638
REFERENCE   2  (bases 1 to 4304)
  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"; doi: 
            "10.1038/nmeth.3147"; journalName: "Nat Methods"; date: "2014-12"; 
            volume: "11"; issue: "12"; pages: "1261-6"
COMMENT     Sequence Label: pReporter_13_switchBP
FEATURES             Location/Qualifiers
     source          1..4304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(295..839)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(375..394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(924..1018)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1039..1698)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1613..1632
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(1861..1890)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     1954..1972
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1976..2019)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2022..2043
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        2044..2078
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     CDS             2106..2783
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2803..2874
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2890..2917
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2924..2944
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     promoter        3100..3134
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(3252..3965)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGNVNGHKFSVSGEGEGDAAYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3292..3312)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     3907..3935
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             3972..3983
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(4014..4063)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      complement(4144..4230)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aggtagagac ccacgaggca gacctcagcg ctagcggagt gtatactggc ttactatgtt
       61 ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg
      121 tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac
      181 gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg
      241 aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca
      301 taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa
      361 cccgacagga ctataaagat accaggcgtt tcccctggcg gctccctcgt gcgctctcct
      421 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
      481 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
      541 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
      601 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
      661 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
      721 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
      781 gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag
      841 aagatcatct tattaagggg tctgacgctc agtggaacga aaagtatata tgagtaaact
      901 tggtctgaca gctcgaggct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
      961 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     1021 agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc
     1081 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
     1141 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa
     1201 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     1261 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     1321 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     1381 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     1441 atcccatatc accagctcac cgtctttcat tgccatacga aattccggat gagcattcat
     1501 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     1561 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     1621 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     1681 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     1741 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat
     1801 caactcgcgc gtttgccacc tgacgtctaa gaaaaggaat attcagcaat ttgcccgtgc
     1861 cgaagaaagg cccacccgtg aaggtgagcc agtgagttga ttgtgccacc tgacgtctaa
     1921 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc
     1981 agataaaaaa aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta
     2041 gagtttacag ctagctcagt cctaggtatt atgctagcta ctagagtcac acaggaaagt
     2101 actagatggc ttcctccgaa gacgttatca aagagttcat gcgtttcaaa gttcgtatgg
     2161 aaggttccgt taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg
     2221 aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg
     2281 acatcctgtc cccgcagttc cagtacggtt ccaaagctta cgttaaacac ccggctgaca
     2341 tcccggacta cctgaaactg tccttcccgg aaggtttcaa atgggaacgt gttatgaact
     2401 tcgaagacgg tggtgttgtt accgttaccc aggactcctc cctgcaagac ggtgagttca
     2461 tctacaaagt taaactgcgt ggtaccaact tcccgtccga cggtccggtt atgcagaaaa
     2521 aaaccatggg ttgggaagct tccaccgaac gtatgtaccc ggaagacggt gctctgaaag
     2581 gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca ctacgacgct gaagttaaaa
     2641 ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac
     2701 tggacatcac ctcccacaac gaagactaca ccatcgttga acagtacgaa cgtgctgaag
     2761 gtcgtcactc caccggtgct taataatact agagccaggc atcaaataaa acgaaaggct
     2821 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     2881 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactagt agcggccgct
     2941 gcagtccggc aaaaaaacgg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
     3001 aagattactt cgcgttatgc aggcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3061 gctgcggcga gcggtatcag ctcactcaaa ggcggtaatt tgacagctag ctcagtccta
     3121 ggtataatgc tagcagttcg atgagagcga taaccaataa cggttgtatt tgtagaactt
     3181 gaccagttgt tttagtaaca taaatacaac tccgaatatt aagcgtagtt ttcgtcgttt
     3241 gctgcaggcc ttttgtatag ttcatccatg ccatgtgtaa tcccagcagc tgttacaaac
     3301 tcaagaagga ccatgtggtc tctcttttcg ttgggatctt tcgaaagggc agattgtgtg
     3361 gacaggtaat ggttgtctgg taaaaggaca gggccatcgc caattggagt attttgttga
     3421 taatggtctg ctagttgaac gcttccatct tcaatgttgt gtctaatttt gaagttaact
     3481 ttgattccat tcttttgttt gtctgccatg atgtatacat tgtgtgagtt atagttgtat
     3541 tccaacttgt gtccaagaat gtttccatct tctttaaaat caataccttt taactcgatt
     3601 ctattaacaa gtgtatcacc ttcaaacttg acttcagcac gtgtcttata gttcccgtca
     3661 tctttgaaaa atatagttct ttcctgtaca taaccttcgg gcatggcact cttgaaaaag
     3721 tcatgccgtt tcatatgatc tgggtatctt gaaaagcatt gaacaccata agtcaaagta
     3781 gtgacaagtg ttggccatgg aacaggtagt tttccagtag tgcaaataaa tttaagggta
     3841 agttttccgt atgctgcatc accttcaccc tctccactga cagaaaattt gtgcccatta
     3901 acattaccat ctaattcaac aagaattggg acaactccag tgaaaagttc ttctccttta
     3961 ctcatctagt atttctcctc tttctctaga ttaaacaaaa ttatttgtag aggctgtttc
     4021 gtcctcacgg actcatcaga ccggaaagca catccggtga cagctactca catggattgc
     4081 ttatatttac aggaccaact ggatctggaa aaacaacaac aatgtatgcg tctagggcgg
     4141 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     4201 gtctttcgac tgagcctttc gttttatttg atgccacgcg taccatggga ctagggccca
     4261 tacccaaaaa aaaaccccgc ccctgacagg gcggggtttt tttt
//
