LOCUS       sequence-98989-e        5108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The tet-responsive reporter pBI-MCS-EGFP (Addgene plasmid 16542) was
            modified to express YFP instead of GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5108)
  AUTHORS   Daringer NM, Dudek RM, Schwarz KA, Leonard JN
  TITLE     Modular Extracellular Sensor Architecture for Engineering Mammalian 
            Cell-based Devices.
  JOURNAL   ACS Synth Biol. 2014 Mar 11.
  PUBMED    24611683
REFERENCE   2  (bases 1 to 5108)
  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: "ACS Synth 
            Biol. 2014 Mar 11."
COMMENT     Sequence Label: pBI-MCS-EYFP
FEATURES             Location/Qualifiers
     source          1..5108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(135..153,154..176,177..195,196..218,219..237,238..260,
                     261..279,280..302,303..321,322..344,345..363,364..386,
                     387..405)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        438..475
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          657..1229
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1361..1380)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1426..1481
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1480..1499)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1872..1889)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2043..2631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2123..2142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2802..3593,3594..3662))
                     /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     3425..3444
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3663..3767)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3835..3853
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    4198..4332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4235..4254)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4289..4308
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4369..5082,5083..5085,5086..5088))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4414..4435)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4763..4782
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 tgcaggaatt ccgggccgcg gaggctggat cggtcccggt gtcttctatg gaggtcaaaa
       61 cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct gcttatatag
      121 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
      181 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
      241 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
      301 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      361 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
      421 ctcggtaccc gggtcgagta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt
      481 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      541 ccgggaccga tccagcctcc gcggccccga attcgagctc ggtacccggg gatcctctag
      601 tcagctgacg cgtgctagcg cggcctcgac gatatctcta gactgagaac ttcagggtga
      661 gtttggggac ccttgattgt tctttctttt tcgctattga aaaattcatg ttatatggag
      721 ggggcaaagt tttcagggtg ttgtttagaa tgggaagatg tcccttgtat caccatggac
      781 cctcatgata attttgtttc tttcactttc tactctgttg acaaccattg tctcctctta
      841 ttttcttttc attttctgta acttttttcg ttaaacttta gcttgcattt gtaacgaatt
      901 tttaaattca ctttcgttta tttgtcagat tgtaagtact ttctctaatc actttttttt
      961 caaggcaatc agggtaatta tattgtactt cagcacagtt ttagagaaca attgttataa
     1021 ttaaatgata aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta
     1081 ttggtagaaa caactacatc ctggtaatca tcctgccttt ctctttatgg ttacaatgat
     1141 atacactgtt tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc
     1201 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt
     1261 ctcatcattt tggcaaagaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg
     1321 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
     1381 attatgggga catcatgaag ccccttgagc atctgacttc tgggtaataa aggaaattta
     1441 ttttcattgc aatagtgtgt gggaattttt tgtgtctctc actcggaagg acatatggga
     1501 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgcca
     1561 tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat gaaacagccc
     1621 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     1681 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     1741 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga
     1801 actcgactgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1861 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1921 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1981 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2041 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2101 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2161 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2221 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2281 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2341 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2401 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2461 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     2521 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2581 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2641 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2701 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2761 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2821 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2881 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2941 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3001 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3061 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3121 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3181 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3241 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3301 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3361 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3421 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3481 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3541 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3601 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3661 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3721 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3781 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3841 cgtatcacga ggccctttcg tcttcactcg acgctggtcg agctgatact tcccgtccgc
     3901 caggggacat gccggcgatg ctgaaggtcg cgcgcattcc cgatgaagag gccggttacc
     3961 gcctgttgac ctggtgggac gggcagggcg ccgcccgagt cttgcctcgg cggcgggcgc
     4021 tctgctcatg gagcgcgcgt ccggggacct tgcacagata gcgtggtccg gccagacgac
     4081 gaggcttgca ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     4141 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     4201 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4261 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4321 catgtctgga tcctctagaa ctaggtcgac aagcttgata tcgaattctt acttgtacag
     4381 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     4441 gcgcttctcg ttggggtctt tgctcagggc ggactggtag ctcaggtagt ggttgtcggg
     4501 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     4561 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
     4621 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     4681 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     4741 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     4801 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     4861 ggggtagcgg gcgaagcact gcaggccgta gccgaaggtg gtcacgaggg tgggccaggg
     4921 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     4981 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     5041 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatac tagttctaga
     5101 gcggccgc
//
