LOCUS       sequence-452336-        5870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian protein expression of a TGF-beta pathway reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  TITLE     Esposito Lab Plasmids 2024
REFERENCE   2  (bases 1 to 5870)
  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"
COMMENT     Sequence Label: 18065-M10-366
FEATURES             Location/Qualifiers
     source          1..5870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    219..239
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1664..1688
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1691..3343
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     protein_bind    complement(3342..3366)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3394..3411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3400..3607
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3649..3665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3649..3665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3662..3684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3673..3689
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3697..3703,3704..3721,3722..3727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3742..3763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3880..3897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4051..4639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4131..4150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4810..5601,5602..5670))
                     /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     5433..5452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5671..5775)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5843..5861
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgga tccgtttaaa cttgaggtac ctcgatcaca actttgtata gaaaagttgg
      241 cgtttatctt cagctggagt cagggaaaca gcgtctcggg aacatcctgt ggactgctag
      301 ctggacagac aagagggtca gtccctccca gggcagggca gacataggct gggggcaaag
      361 taaacatgcc ccagggtggc tagactgcgg gagatctggt cgttcagtca ctccaccagt
      421 agctgatcag tagataactt gaggtgaggc tcagatctgg aaacagcgtc tcgggaacat
      481 cctgtggact gctagctgga cagacaagag ggtcagtccc tcccagggca gggcagacat
      541 aggctggggg caaagtaaac atgccccagg gtggctagac tgcggggaat tccactacag
      601 gccgggtatt taaaaccgcg ggcccaataa gcggcatcat atgaaatgaa ttcggaaaca
      661 gcgtctcggg aacatcctgt ggactgctag ctggacagac aagagggtca gtccctccca
      721 gggcagggca gacataggct gggggcaaag taaacatgcc ccagggtggc tagactgcgg
      781 gccatgggcc gcgcggaccc tcccatgacg tatgcatcgt atgctagtgc tgttatggag
      841 tgccatgggg aaacagcgtc tcgggaacat cctgtggact gctagctgga cagacaagag
      901 ggtcagtccc tcccagggca gggcagacat aggctggggg caaagtaaac atgccccagg
      961 gtggctagac tgcgggtcgc gaattgtgcg cactttaccg gcggggacac catgaagtgt
     1021 gcttctgctg accatcctcg cgaggaaaca gcgtctcggg aacatcctgt ggactgctag
     1081 ctggacagac aagagggtca gtccctccca gggcagggca gacataggct gggggcaaag
     1141 taaacatgcc ccagggtggc tagactgcgg ggtttaaaca ctgcccgccg aaaaatggat
     1201 aagcatagta agctgcataa gtggacaaca ctcagtttaa acggaaacag cgtctcggga
     1261 acatcctgtg gactgctagc tggacagaca agagggtcag tccctcccag ggcagggcag
     1321 acataggctg ggggcaaagt aaacatgccc cagggtggct agactgcggg tcgaggaagc
     1381 ccctccccct gcagccgcgc agagccaccc ggctcgtggc cgctgtttac aaggacacgc
     1441 gcttcctgac agtgacgcga gccgcctcct ccccttcccc acgctctagg agggggccgc
     1501 gggggcctgg ctcccgcgtc ggccaatcgg agtgcacttc cgcagctgac aaattcagta
     1561 taaaagcttg gggctggggc cgagcactgg ggactttgag ggtggccagg ccagcgtagg
     1621 atcctgctgg gagcggggaa ctgagggaag cgacgccgat aaaacaactt tgtacaaaaa
     1681 agttggcacc atggaagatg ccaaaaacat taagaagggc ccagcgccat tctacccact
     1741 cgaagacggg accgccggcg agcagctgca caaagccatg aagcgctacg ccctggtgcc
     1801 cggcaccatc gcctttaccg acgcacatat cgaggtggac attacctacg ccgagtactt
     1861 cgagatgagc gttcggctgg cagaagctat gaagcgctat gggctgaata caaaccatcg
     1921 gatcgtggtg tgcagcgaga atagcttgca gttcttcatg cccgtgttgg gtgccctgtt
     1981 catcggtgtg gctgtggccc cagctaacga catctacaac gagcgcgagc tgctgaacag
     2041 catgggcatc agccagccca ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct
     2101 caacgtgcaa aagaagctac cgatcataca aaagatcatc atcatggata gcaagaccga
     2161 ctaccagggc ttccaaagca tgtacacctt cgtgacttcc catttgccac ccggcttcaa
     2221 cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa accatcgccc tgatcatgaa
     2281 cagtagtggc agtaccggat tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt
     2341 ccgattcagt catgcccgcg accccatctt cggcaaccag atcatccccg acaccgctat
     2401 cctcagcgtg gtgccatttc accacggctt cggcatgttc accacgctgg gctacttgat
     2461 ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag gagctattct tgcgcagctt
     2521 gcaagactat aagattcaat ctgccctgct ggtgcccaca ctatttagct tcttcgctaa
     2581 gagcactctc atcgacaagt acgacctaag caacttgcac gagatcgcca gcggcggggc
     2641 gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc ttccacctac caggcatccg
     2701 ccagggctac ggcctgacag aaacaaccag cgccattctg atcacccccg aaggggacga
     2761 caagcctggc gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg tggacttgga
     2821 caccggtaag acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat
     2881 catgagcggc tacgttaaca accccgaggc tacaaacgct ctcatcgaca aggacggctg
     2941 gctgcacagc ggcgacatcg cctactggga cgaggacgag cacttcttca tcgtggaccg
     3001 gctgaagagc ctgatcaaat acaagggcta ccaggtagcc ccagccgaac tggagagcat
     3061 cctgctgcaa caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc
     3121 cggcgagctg cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga
     3181 gatcgtggac tatgtggcca gccaggttac aaccgccaag aagctgcgcg gtggtgttgt
     3241 gttcgtggac gaggtgccta aaggactgac cggcaagttg gacgcccgca agatccgcga
     3301 gattctcatt aaggccaaga agggcggcaa gatcgccgtg taaccaactt tcttgtacaa
     3361 agttgtgatg accttctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     3421 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3481 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3541 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3601 ctggggagat ctgtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     3661 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3721 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3781 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3841 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3901 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3961 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4861 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4921 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4981 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5041 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5101 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5161 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5221 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5281 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5341 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5401 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5461 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5521 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5581 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5641 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5701 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5761 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5821 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
