LOCUS       sequence-125537-        4592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP from J23101 promoter and CFP from standard J23101 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4592)
  AUTHORS   Grant PK, Dalchau N, Brown JR, Federici F, Rudge TJ, Yordanov B, 
            Patange O, Phillips A, Haseloff J
  TITLE     Orthogonal intercellular signaling for programmed spatial behavior.
  JOURNAL   Mol Syst Biol. 2016 Jan 25;12(1):849.
  PUBMED    26814193
REFERENCE   2  (bases 1 to 4592)
  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: "Mol Syst 
            Biol."; date: "2016-01-25"; volume: "12(1)"; pages: "849"
COMMENT     Sequence Label: pJ23101
FEATURES             Location/Qualifiers
     source          1..4592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             44..55
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(62..64,65..67,68..781)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      801..872
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      888..915
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    922..942
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(1160..1975)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1160..1975)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1883..1902
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1967..1986
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      complement(2135..2162)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2178..2249)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(2269..2982,2983..2985,2986..2988))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     RBS             2995..3006
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3015..3049)
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     rep_origin      complement(3491..4035)
                     /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(3571..3590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4503..4521
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4525..4568)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4571..4592
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tttacagcta gctcagtcct aggtattatg ctagctacta gagaaagagg agaaatacta
       61 gatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      121 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      181 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      241 cctcgtgacc accttcggct acggcctgca atgcttcgcc cgctaccccg accacatgaa
      301 gctgcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      361 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      421 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      481 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
      541 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
      601 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
      661 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
      721 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
      781 ataatactag agccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
      841 gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg
      901 gtgggccttt ctgcgtttat atactagtag cggccgctgc agtccggcaa aaaaacgggc
      961 aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg cgttatgcag
     1021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1081 cactcaaagg cggtaatctc gagtcccgtc aagtcagcgt aatgctctgc cagtgttaca
     1141 accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     1201 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     1261 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     1321 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     1381 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc
     1441 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     1501 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     1561 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1621 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1681 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1741 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1801 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1861 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1921 tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg ctcataacac
     1981 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     2041 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa
     2101 cttttgctga gttgaaggat cagatcacta taaacgcaga aaggcccacc cgaaggtgag
     2161 ccagtgtgac tctagtagag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     2221 tctttcgact gagcctttcg ttttatttga tgcctggctc tagtattatt acttgtacag
     2281 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     2341 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg
     2401 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     2461 gctgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt tcttctgctt
     2521 gtcggcggtg atatagacgt tgtggctgat gtagttgtac tccagcttgt gccccaggat
     2581 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     2641 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     2701 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     2761 ggggtagcgg ctgaagcact gcacgcccca ggtcagggtg gtcacgaggg tgggccaggg
     2821 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     2881 gccctcgccc tcgccggaca cgctgaactt gtggccgttc acgtcgccgt ccagctcgac
     2941 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatct agtatttctc
     3001 ctctttctct agtagctagc ataataccta ggactgagct agctgtaaag catcttcccg
     3061 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     3121 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     3181 cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcagcgctag cggagtgtat
     3241 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     3301 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     3361 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     3421 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     3481 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     3541 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc ctggcggctc
     3601 cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc
     3661 gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg
     3721 actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
     3781 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
     3841 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
     3901 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
     3961 ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
     4021 ccaaaacgat ctcaagaaga tcatcttatt aaggggtctg acgctcagtg gaacgaaaac
     4081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4441 tctattccat ggtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4501 aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg
     4561 atgatttctg gaattcgcgg ccgcttctag ag
//
