LOCUS       sequence-486408-        4075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SP6-GFP plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4075)
  AUTHORS   Kucinski J, Tallan A, Taslim C, Vontell AM, Silvius KM, Wang M, 
            Cannon MV, Stanton BZ, Kendall GC
  TITLE     Rhabdomyosarcoma fusion oncoprotein initially pioneers a neural 
            signature in vivo.
  JOURNAL   Cell Rep. 2025 Jul 22;44(7):115923. doi: 
            10.1016/j.celrep.2025.115923. Epub 2025 Jul 3.
  PUBMED    40614256
REFERENCE   2  (bases 1 to 4075)
  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: "Cell Rep.";
            date: "2025-07-22"; volume: "44(7):115923. doi"; pages: " 
            10.1016/j.celrep.2025.115923. Epub 2025 Jul 3"
COMMENT     Sequence Label: CNTL-sfGFP
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..30
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(249..353)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(431..886)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(568..589)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     780..799
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1012..1034
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1026..1043
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1027..1043
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1050..1068
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1099..1108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1103..1112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1128..1144
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1163..1183
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1165..1183
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1296..1430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1333..1352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1387..1406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(1460..2173,2174..2176))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(1463..2176)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1503..1523)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2118..2146
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        complement(2266..2284)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2267..2284)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2302..2318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2315..2337)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2326..2342
                     /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(2350..2356,2357..2374,2375..2380))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2395..2416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2533..2550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2704..3292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2784..2803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3463..4075,1..179,180..248))
                     /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"
ORIGIN
        1 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
       61 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
      121 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      181 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      241 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      361 ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
      421 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      481 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      541 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      601 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      661 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      721 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      781 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      841 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca
      901 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
      961 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     1021 gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggcc
     1081 cgacgtcgca tgctcccggc cgccatggcg gccgcgggaa ttcgattcag gaaacagcta
     1141 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctgggtac
     1201 cgggcccaat gcattggcgc cgcggccgcg aattaaaaaa cctcccacac ctccccctga
     1261 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     1321 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1381 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tacgtaatac
     1441 gactcactat agttctagat tatttgtaga gctcatccat gccatgtgta atcccagcag
     1501 cagttacaaa ctcaagaagg accatgtggt cacgcttttc gttgggatct ttcgacagga
     1561 cagattgtgt cgacaggtaa tggttgtctg gtaaaaggac agggccatcg ccaattggag
     1621 tattttgttg ataatggtct gctagttgaa cggaaccatc ttcaacgttg tggcgaattt
     1681 tgaagttagc tttgattcca ttcttttgtt tgtctgccgt gatgtataca ttgtgtgagt
     1741 taaagttgta ctcgagtttg tgtccaagaa tgtttccatc ttctttaaaa tcaataccct
     1801 ttaactcgat acgattaaca agggtatcac cttcaaactt gacttcagca cgcgtcttgt
     1861 aggtcccgtc atctttgaaa gatatagtgc gttcctgtac ataaccttcg ggcatggcac
     1921 tcttgaaaaa gtcatgccgt ttcatgtgat ccggataacg ggaaaagcat tgaacaccat
     1981 aggtcagagt agtgacaagt gttggccacg gaacaggtag ttttccagta gtgcaaataa
     2041 atttaagggt gagttttccg tttgtagcat caccttcacc ctctccacgg acagaaaatt
     2101 tgtgcccatt aacatcacca tctaattcaa caagaattgg gacaactcca gtgaaaagtt
     2161 cttctccttt gctcatggtg gcggcaatca ctagtgaatt cgcggccgcc tgcaggtcga
     2221 ccatatggga gagctcccaa cgcgttggat gcatagcttg agtattctat agtgtcacct
     2281 aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcg
//
