LOCUS       sequence-208497-        4591 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 4591)
  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 4591)
  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..4591
                     /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"
     terminator      943..1000
                     /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."
     CDS             complement(1158..1973)
                     /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(1158..1973)
                     /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     1881..1900
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2133..2160)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2176..2247)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(2267..2980,2981..2983,2984..2986))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     RBS             2993..3004
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3013..3047)
                     /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(3489..4034)
                     /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(3569..3588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4502..4520
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4524..4567)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4570..4591
                     /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 aaaagggcaa
      961 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc
     1021 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1081 ctcaaaggcg gtaatctcga gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac
     1141 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     1201 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     1261 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     1321 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     1381 tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat ttccttccag
     1441 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     1501 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     1561 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     1621 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     1681 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     1741 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     1801 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     1861 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     1921 ttggaattta atcgcggcct ggagcaagac gtttcccgtt gaatatggct cataacacca
     1981 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     2041 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact
     2101 tttgctgagt tgaaggatca gatcactata aacgcagaaa ggcccacccg aaggtgagcc
     2161 agtgtgactc tagtagagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     2221 tttcgactga gcctttcgtt ttatttgatg cctggctcta gtattattac ttgtacagct
     2281 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     2341 gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg ttgtcgggca
     2401 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     2461 tgccgtcctc gatgttgtgg cggatcttga agttggcctt gatgccgttc ttctgcttgt
     2521 cggcggtgat atagacgttg tggctgatgt agttgtactc cagcttgtgc cccaggatgt
     2581 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     2641 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     2701 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     2761 ggtagcggct gaagcactgc acgccccagg tcagggtggt cacgagggtg ggccagggca
     2821 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
     2881 cctcgccctc gccggacacg ctgaacttgt ggccgttcac gtcgccgtcc agctcgacca
     2941 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatctag tatttctcct
     3001 ctttctctag tagctagcat aatacctagg actgagctag ctgtaaagca tcttcccgac
     3061 aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac
     3121 aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcgggcc
     3181 tggtatgagt cagcaacacc ttcttcacga ggcagacctc agcgctagcg gagtgtatac
     3241 tggcttacta tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa
     3301 aaaggctgca ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct
     3361 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg
     3421 cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca
     3481 aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat
     3541 cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc
     3601 ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg
     3661 cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga
     3721 ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct
     3781 tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt
     3841 tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg
     3901 gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct
     3961 tcgaaaaacc gccctgcaag gcggtttttt cgttctcaga gcaagagatt acgcgcagac
     4021 caaaacgatc tcaagaagat catcttatta aggggtctga cgctcagtgg aacgaaaact
     4081 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4141 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4201 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4261 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4321 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4381 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4441 ctattccatg gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4501 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
     4561 tgatttctgg aattcgcggc cgcttctaga g
//
