LOCUS       sequence-447363-        5181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23110_buj_sfGFP_buj_mRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5181)
  AUTHORS   Cardiff RAL, Faulkner ID, Beall JG, Carothers JM, Zalatan JG
  TITLE     
  JOURNAL   Nucleic Acids Res. 2024 May 22;52(9):5406-5419. doi: 
            10.1093/nar/gkae275.
  PUBMED    38613390
REFERENCE   2  (bases 1 to 5181)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2024-05-22- 22"; volume: "52"; issue: 
            "9"; pages: "5406-5419"
COMMENT     Sequence Label: pRC072
FEATURES             Location/Qualifiers
     source          1..5181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(249..343)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(373..1164,1165..1233))
                     /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"
     primer_bind     996..1015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1234..1338)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1551..1585
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     RBS             1595..1606
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1613..1615,1616..2329)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1613..2326
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1643..1671)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2266..2286
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     RBS             2345..2356
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2363..3040
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3073..3144
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3160..3187
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3718..4668)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
       61 ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc
      121 tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg
      181 gctaatgcac ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcccta
      241 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
      301 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgtaaac
      361 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
      421 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
      481 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
      541 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
      601 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
      661 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
      721 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
      781 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
      841 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
      901 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
      961 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1021 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1081 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1141 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1201 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     1261 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1321 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgtggcaa ttccgacgtc
     1381 agcatttgcg atcattcacg cagcgcttat tcagttgctc actgcgatgt cataatcatc
     1441 gctacgagct gtgaaagatg cataaagctc gtacgacgcg ttcgctcgtc tcctcacttc
     1501 tcctacggag cgttctggac acaacgtcgt cttgaagttg cgattataga tttacggcta
     1561 gctcagtcct aggtacaatg ctagcgaatt cattaaagag gagaaaggta ccatgagcaa
     1621 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     1681 tgggcacaaa ttttctgtcc gtggagaggg tgaaggtgat gctacaaacg gaaaactcac
     1741 ccttaaattt atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac
     1801 tctgacctat ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt
     1861 tttcaagagt gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga
     1921 cgggacctac aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat
     1981 cgagttaaag ggtattgatt ttaaagaaga tggaaacatt cttggacaca aactcgagta
     2041 caactttaac tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc
     2101 taacttcaaa attcgccaca acgttgaaga tggttccgtt caactagcag accattatca
     2161 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtcgac
     2221 acaatctgtc ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt
     2281 tgtaactgct gctgggatta cacatggcat ggatgagctc tacaaataag gatccgaatt
     2341 cattaaagag gagaaaggta ccatggcgag tagcgaagac gttatcaaag agttcatgcg
     2401 tttcaaagtt cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg
     2461 tgaaggtcgt ccgtacgaag gtacccagac cgctaaactg aaagttacca aaggtggtcc
     2521 gctgccgttc gcttgggaca tcctgtcccc gcagttccag tacggttcca aagcttacgt
     2581 taaacacccg gctgacatcc cggactacct gaaactgtcc ttcccggaag gtttcaaatg
     2641 ggaacgtgtt atgaacttcg aagacggtgg tgttgttacc gttacccagg actcctccct
     2701 gcaagacggt gagttcatct acaaagttaa actgcgtggt accaacttcc cgtccgacgg
     2761 tccggttatg cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta tgtacccgga
     2821 agacggtgct ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta
     2881 cgacgctgaa gttaaaacca cctacatggc taaaaaaccg gttcagctgc cgggtgctta
     2941 caaaaccgac atcaaactgg acatcacctc ccacaacgaa gactacacca tcgttgaaca
     3001 gtacgaacgt gctgaaggtc gtcactccac cggtgcttaa ggatccaaac tcgagtaagg
     3061 atctccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     3121 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     3181 ttctgcgttt atacctaggg tacgggtttt gctgcccgca aacgggctgt tctggtgttg
     3241 ctagtttgtt atcagaatcg cagatccggc ttcagccggt ttgccggctg aaagcgctat
     3301 ttcttccaga attgccatga ttttttcccc acgggaggcg tcactggctc ccgtgttgtc
     3361 ggcagctttg attcgataag cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga
     3421 gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta gttgctttgt tttactggtt
     3481 tcacctgttc tattaggtgt tacatgctgt tcatctgtta cattgtcgat ctgttcatgg
     3541 tgaacagctt tgaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca
     3601 ccgttttcat ctgtgcatat ggacagtttt ccctttgata tgtaacggtg aacagttgtt
     3661 ctacttttgt ttgttagtct tgatgcttca ctgatagata caagagccat aagaacctca
     3721 gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca
     3781 tgagaacgaa ccattgagat catacttact ttgcatgtca ctcaaaaatt ttgcctcaaa
     3841 actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttatg
     3901 taggtaggaa tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt
     3961 gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt
     4021 atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc
     4081 tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc
     4141 aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt
     4201 cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa
     4261 agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa
     4321 tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca
     4381 ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag
     4441 ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg
     4501 agcgtattgg ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt
     4561 ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata
     4621 gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg
     4681 gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt
     4741 ccttttcccg ggtgatctgg gtatctgtaa attctgctag acctttgctg gaaaacttgt
     4801 aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat
     4861 tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc
     4921 cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa ggatgtcgca
     4981 aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct
     5041 cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc
     5101 gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta
     5161 aatggcacta caggcgcctt t
//
