LOCUS       sequence-107122-        4335 bp    DNA     circular SYN 15-APR-2025
DEFINITION  A reporter plasmid containing the attB and attP sites flanking a 
            green fluorescent protein reporter gene (gfp)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4335)
  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 4335)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_10
FEATURES             Location/Qualifiers
     source          1..4335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(295..840)
                     /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(925..1019)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1040..1699)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1614..1633
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(1862..1891)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     1955..1973
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1977..2020)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2023..2044
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        2045..2079
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     CDS             2107..2784
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2804..2875
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2891..2918
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2946..3003
                     /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."
     promoter        3099..3133
                     /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(4175..4261)
                     /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 tccccctggc ggctccctcg tgcgctctcc
      421 tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc
      481 cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac
      541 cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      601 aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc
      661 ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct
      721 ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc
      781 tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa
      841 gaagatcatc ttattaaggg gtctgacgct cagtggaacg aaaagtatat atgagtaaac
      901 ttggtctgac agctcgaggc ttggattctc accaataaaa aacgcccggc ggcaaccgag
      961 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     1021 aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     1081 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     1141 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     1201 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     1261 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     1321 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     1381 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     1441 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
     1501 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     1561 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     1621 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     1681 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     1741 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga
     1801 tcaactcgcg cgtttgccac ctgacgtcta agaaaaggaa tattcagcaa tttgcccgtg
     1861 ccgaagaaag gcccacccgt gaaggtgagc cagtgagttg attgtgccac ctgacgtcta
     1921 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt
     1981 cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcgc ggccgcttct
     2041 agagtttaca gctagctcag tcctaggtat tatgctagct actagagtca cacaggaaag
     2101 tactagatgg cttcctccga agacgttatc aaagagttca tgcgtttcaa agttcgtatg
     2161 gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac
     2221 gaaggtaccc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg
     2281 gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca cccggctgac
     2341 atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg tgttatgaac
     2401 ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga cggtgagttc
     2461 atctacaaag ttaaactgcg tggtaccaac ttcccgtccg acggtccggt tatgcagaaa
     2521 aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg tgctctgaaa
     2581 ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc tgaagttaaa
     2641 accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac cgacatcaaa
     2701 ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga acgtgctgaa
     2761 ggtcgtcact ccaccggtgc ttaataatac tagagccagg catcaaataa aacgaaaggc
     2821 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     2881 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatatactag tagcggcagc
     2941 tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa
     3001 agattacttc gcgttatgca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3061 ctgcggcgag cggtatcagc tcactcaaag gcggtaattt gacagctagc tcagtcctag
     3121 gtataatgct agcagttcga tgagagcgat aacaagcacg ctgataatca gcaagaccac
     3181 caacatttcc accaatgtaa aagctttaac cttagctttt 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 cagctaaaag agtgagagtt
     4081 ttactatcct tgattgagat gtaggttact aaaattattt atattttcca aacagaggac
     4141 acatgcccta cctccatgat ctctagggcg gcggatttgt cctactcagg agagcgttca
     4201 ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt
     4261 gatgccacgc gtaccatggg actagggccc atacccaaaa aaaaaccccg cccctgacag
     4321 ggcggggttt ttttt
//
