LOCUS       sequence-211335-        7042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  reporter of STAT3 signaling activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7042)
  AUTHORS   Wei W, Tweardy DJ, Zhang M, Zhang X, Landua J, Petrovic I, Bu W, 
            Roarty K, Hilsenbeck SG, Rosen JM, Lewis MT
  TITLE     STAT3 signaling is activated preferentially in tumor-initiating 
            cells in claudin-low models of human breast cancer.
  JOURNAL   Stem Cells. 2014 Oct;32(10):2571-82. doi: 10.1002/stem.1752.
  PUBMED    24891218
REFERENCE   2  (bases 1 to 7042)
  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"; doi: "10.1002/stem"; 
            journalName: "Stem Cells"; date: "2014-10"; volume: "32"; issue: 
            "10"; pages: "2571-82"
COMMENT     Sequence Label: STAT3-GFP
FEATURES             Location/Qualifiers
     source          1..7042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    254..371
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      588..597
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(594..596,597..599,600..1313)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(594..596,597..599,600..1310)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(639..660)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(900..919)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1247..1268
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1338..1926
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1391..1411)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1809..1820)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2013..2246
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2318..2439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2355..2374)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2409..2428
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2479..2614
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2634..2653)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2635..2653)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2663..2680)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2663..2679)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2672..2694)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2821..3276
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2908..2927)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3118..3139
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3302..3406
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3407..3475,3476..4267)
                     /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(3625..3644)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4438..5026
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4927..4946
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5180..5197
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5314..5335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5350..5355,5356..5373,5374..5380)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5388..5404
                     /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     5393..5415
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5412..5428
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5412..5428
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5447..5467
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5449..5467
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5495..5721
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5722..5902
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5949..6074
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6567..6800
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6985..7029
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
       61 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
      121 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
      181 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
      241 ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
      301 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
      361 ttcaaaattt tatcgatcac gagactagcc tcctagcccg ggctcgagca gctgaagctt
      421 gcatgccctg cagggttccc gtaaatgcat caggttcccg taaatgcatc aggttcccgt
      481 aaatgcatca ggttcccgta aatgcatcac tgcagagggt atataatgga tccggtactc
      541 gagatctgcg atctaagtaa gcttggcatt ccggtactgt tggtaaagcc accatggtga
      601 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      661 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      721 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      781 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
      841 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      901 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      961 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1021 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1081 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1141 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1201 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1261 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     1321 gcgactctag agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1381 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1441 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     1501 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     1561 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     1621 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     1681 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     1741 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     1801 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     1861 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     1921 ccccgcctgg aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga
     1981 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     2041 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     2101 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     2161 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     2221 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     2281 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     2341 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2401 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     2461 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2521 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2581 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     2641 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2701 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2761 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     2821 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2881 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2941 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3001 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3061 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3121 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3181 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3241 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     3301 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3361 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3421 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3481 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3541 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3601 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3661 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3721 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3781 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3841 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3901 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3961 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4021 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4081 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4141 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4201 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4261 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4321 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4381 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4441 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4501 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4561 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4621 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4681 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4741 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4801 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4861 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4921 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4981 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5041 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5101 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5161 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     5221 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     5281 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     5341 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     5401 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     5461 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
     5521 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     5581 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     5641 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     5701 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     5761 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     5821 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     5881 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     5941 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     6001 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     6061 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     6121 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     6181 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     6241 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     6301 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     6361 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     6421 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     6481 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     6541 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     6601 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     6661 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     6721 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     6781 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     6841 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     6901 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     6961 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     7021 gcaagtttgt ggaattggtt ta
//
