LOCUS       sequence-435942-        5701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TGFb-inducible fluorescent reporter (TIE:EGFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  TITLE     Tgfb-Induced-Enhancer:Beta-globin-minimal-promoter-EGFP:pA 
            pDestTol2pA2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5701)
  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: 
            "Unpublished"
COMMENT     Sequence Label: 
            Tgfb-Induced-Enhancer:Beta-globin-minimal-promoter-EGFP:pA 
            pDestTol2pA2
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     473..490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    607..628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(643..648,649..666,667..673)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    681..697
                     /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)."
     primer_bind     686..708
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     705..721
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     740..760
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        742..760
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     797..813
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     797..813
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(847..858,859..1046)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    1485..1619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1635..1651
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1659..1679
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    1742..1876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1878..1902
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1964..2674,2675..2677,2678..2680))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1964..2674,2675..2677,2678..2680))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2006..2027)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2355..2374
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2614..2635
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2821..2845)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3160..3180)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(3200..3216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(3589..3751,3752..3763)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     4112..4133
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4296..4400
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4401..4469,4470..5261)
                     /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     complement(4619..4638)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5432..5701,1..319)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
       61 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      481 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      541 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      601 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      661 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      721 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctcca
      781 ccgcggtggc ggccgctcta gaactagtgg atctgctggg cttgctgaag gtagggggtc
      841 aagaaccaga ggtgtaaagt acttgagtaa ttttacttga ttactgtact taagtattat
      901 ttttggggat ttttacttta cttgagtaca attaaaaatc aatactttta cttttactta
      961 attacatttt tttagaaaaa aaagtacttt ttactcctta caattttatt tacagtcaaa
     1021 aagtacttat tttttggaga tcacttcatt ctattttccc ttgctattac caaaccaatt
     1081 gaattgcgct gatgcccagt ttaatttaaa tgttatttat tctgcctatg aaaatcgttt
     1141 tcacattata tgaaattggt cagacatgtt cattggtcct ttggaagtga cgtcatgtca
     1201 catctattac cacaatgcac agcaccttga cctggaaatt agggaaatta taacagtcaa
     1261 tcagtggaag aaaatggagg aagtatgtga ttcatcagca gctgcgagca gcacagtcca
     1321 aaatcagcca caggatcaag agcacccgtg gccgtatctt cgcagatctg atctagagga
     1381 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     1441 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     1501 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1561 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
     1621 tcatcgatgg taccgtaaaa cgacggccag tgaattatca actatgtata ataaagttgg
     1681 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     1741 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1801 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1861 ttatcatgtc tggatccccc actttgtaca agaaagctgg gtgctatagg gctgcagaat
     1921 ctagaggctc gagaggcctt gaattcccag atcttccacc gcccttgtac agctcgtcca
     1981 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     2041 cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca
     2101 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     2161 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     2221 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     2281 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     2341 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     2401 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     2461 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca
     2521 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     2581 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     2641 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat gggatgtctg tttctgaggt
     2701 tgcaagtcaa cacaacagac tcagaagcaa acgtaagaag caactgaccc tgctcagctt
     2761 tatataccca atgctggctc ctgcccactc tgtcctctct gagcagattg gccatggtcc
     2821 agcctgcttt tttgtacaaa cttgggtcga attcgccctt gacaggtcgc acctgagtcc
     2881 ccaaacagca gaggaacccc gacggctcgc ttgcctgctc tcctgcaggc cgcccctgct
     2941 ggcagaggtc cctgatgact gggggtctgg caatgccagc tcacagacca catgccaccc
     3001 caaggtgttt gctttgctca gctgggtggg tgagtcatct cagagccccc ctcccttgcc
     3061 tgtggctgtg caagggcttt ggcagctggg atttgaggaa tctcaagccc ctctcgattt
     3121 gctctaccag gatgacaaag aaaagggcga attcggagcc aacttttcta tacaaagttg
     3181 atagcttggc gtaatcatgg tcatagctgt ttcctgctcg agggcccatc tggcctgtgt
     3241 ttcagacacc agggagtctc tgctcacgtt tcctgctatt tgcagcctct ctatcaagac
     3301 taatacacct cttcccgcat cggctgcctg tgagaggctt ttcagcactg caggattgct
     3361 tttcagcccc aaaagagcta ggcttgacac taacaatttt gagaatcagc ttctactgaa
     3421 gttaaatctg aggttttaca actttgagta gcgtgtactg gcattagatt gtctgtctta
     3481 tagtttgata attaaataca aacagttcta aagcaggata aaaccttgta tgcatttcat
     3541 ttaatgtttt ttgagattaa aagcttaaac aagaatctct agttttcttt cttgctttta
     3601 cttttacttc cttaatactc aagtacaatt ttaatggagt acttttttac ttttactcaa
     3661 gtaagattct agccagatac ttttactttt aattgagtaa aattttccct aagtacttgt
     3721 actttcactt gagtaaaatt tttgagtact ttttacacct ctgtcaagaa ctcctggaca
     3781 aacctctgac ctgtgtggaa cagagtggat atgggtgtct gaacagatat tcacgtcttt
     3841 tgcagatcag agggcatttc tggtgttgtg caatgacctg ggtccaacag gtaggcgtac
     3901 tcgtatccaa gagcattgtt ggacaggagg tagtctccat tcctataaag aggaaggaat
     3961 ggcaccatgt agtagccgtc attgtggcca attggtgcat tggtacgtgg gtagatggac
     4021 cggggaggct gatgagttct tagccagcgt gggccatcgc cctgatagac ggtttttcgc
     4081 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4141 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4201 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4261 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4321 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4381 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4441 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4501 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4561 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4621 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4681 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4741 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4801 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4861 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4921 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4981 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5041 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5101 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5161 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5221 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5281 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5341 tcctttttga taatctcatg accaaaatgc cttaacgtga gttttcgttc cactgagcgt
     5401 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5461 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5521 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5581 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5641 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5701 g
//
