LOCUS       sequence-211331-        6960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  negative control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  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 6960)
  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: TATA-GFP
FEATURES             Location/Qualifiers
     source          1..6960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    233..350
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      485..494
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(491..493,494..496,497..1210)
                     /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(491..493,494..496,497..1207)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(536..557)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(797..816)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1144..1165
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1235..1823
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1288..1308)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1706..1717)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1910..2143
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2215..2336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2252..2271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2306..2325
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2376..2511
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2531..2550)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2532..2550)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2560..2577)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2560..2576)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2569..2591)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2718..3173
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2805..2824)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3015..3036
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3199..3303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3304..3372,3373..4164)
                     /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(3522..3541)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4335..4923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4824..4843
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5077..5094
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5211..5232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5247..5252,5253..5270,5271..5277)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5285..5301
                     /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     5290..5312
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5309..5325
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5309..5325
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5344..5364
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5346..5364
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5392..5618
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5619..5799
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5846..5971
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6464..6697
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6882..6926
                     /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 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
       61 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
      121 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
      181 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta acttttaaaa
      241 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
      301 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt atcgatcacg
      361 agactagcct cctagcccgg gctcgagcag ctgaagcttg catgccctgc agagggtata
      421 taatggatcc ggtactcgag atctgcgatc taagtaagct tggcattccg gtactgttgg
      481 taaagccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      541 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      601 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      661 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      721 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      781 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      841 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      901 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
      961 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1021 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1081 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1141 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1201 gtacaagtaa agcggccgcg actctagagt cgacaatcaa cctctggatt acaaaatttg
     1261 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1321 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1381 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1441 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     1501 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     1561 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     1621 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     1681 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     1741 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     1801 ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacctttaag accaatgact
     1861 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     1921 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     1981 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     2041 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     2101 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     2161 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     2221 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2281 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2341 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2401 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2461 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     2521 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     2581 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     2641 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2701 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2761 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2821 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2881 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2941 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3001 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3061 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3121 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3181 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3241 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3301 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3361 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3421 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3481 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3541 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3601 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3661 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3721 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3781 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3841 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3901 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3961 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4021 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4081 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4141 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4201 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4261 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4321 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4381 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4441 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4501 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4561 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4621 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4681 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4741 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4801 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4861 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4921 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4981 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5041 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5101 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5161 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5221 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5281 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     5341 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     5401 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     5461 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     5521 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     5581 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     5641 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     5701 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     5761 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     5821 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     5881 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     5941 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     6001 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     6061 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     6121 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     6181 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     6241 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     6301 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     6361 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     6421 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     6481 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     6541 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     6601 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     6661 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     6721 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     6781 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     6841 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     6901 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
//
