LOCUS       sequence-120988-        4928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF8 Op -- GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4928)
  AUTHORS   Slomovic S, Collins JJ
  TITLE     DNA sense-and-respond protein modules for mammalian cells.
  JOURNAL   Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585.
  PUBMED    26389572
REFERENCE   2  (bases 1 to 4928)
  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: "Nat 
            Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585."
COMMENT     Sequence Label: pGL4.26-SS-139
FEATURES             Location/Qualifiers
     source          1..4928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..87
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      114..123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(120..122,123..125,126..839)
                     /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(120..122,123..125,126..836)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(165..186)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(426..445)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     773..794
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    883..1004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(894..913)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     948..967
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1198..1555
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1198..1389
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1221..1241)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1406..1541
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1468..1487
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1586..2623
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2602..2610
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    2647..2695
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2851..2868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3022..3610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3102..3121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3810..4601,4602..4670))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4775..4823
                     /note="synthetic polyadenylation signal"
     misc_feature    4837..4928
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4877..4896
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gccgaagtgg tccaagctta gacactagag
       61 ggtatataat ggaagctcga cttccagctt ggcaatccgg tactgttggt aaagccacca
      121 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      181 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      241 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      301 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
      361 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
      421 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      481 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
      541 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
      601 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
      661 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
      721 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
      781 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
      841 aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt
      901 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
      961 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1021 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     1081 tctacaaatg tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc ttccgctgat
     1141 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag ctaccttctg
     1201 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     1261 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     1321 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1381 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     1441 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     1501 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     1561 cgattcttct gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga
     1621 aaaatttctc atcgagaagt tcgacagtgt gagcgacctg atgcagttgt cggagggcga
     1681 agagagccga gccttcagct tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag
     1741 ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact
     1801 acccatcccc gaagtgttgg acatcggcga gttcagcgag agcctgacat actgcatcag
     1861 tagacgcgcc caaggcgtta ctctccaaga cctccccgaa acagagctgc ctgctgtgtt
     1921 acagcctgtc gccgaagcta tggatgctat tgccgccgcc gacctcagtc aaaccagcgg
     1981 cttcggccca ttcgggcccc aaggcatcgg ccagtacaca acctggcggg atttcatttg
     2041 cgccattgct gatccccatg tctaccactg gcagaccgtg atggacgaca ccgtgtccgc
     2101 cagcgtagct caagccctgg acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg
     2161 ccacctcgtc catgccgact tcggcagcaa caacgtcctg accgacaacg gccgcatcac
     2221 cgccgtaatc gactggtccg aagctatgtt cggggacagt cagtacgagg tggccaacat
     2281 cttcttctgg cggccctggc tggcttgcat ggagcagcag actcgctact tcgagcgccg
     2341 gcatcccgag ctggccggca gccctcgtct gcgagcctac atgctgcgca tcggcctgga
     2401 tcagctctac cagagcctcg tggacggcaa cttcgacgat gctgcctggg ctcaaggccg
     2461 ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt cgcacacaaa tcgctcgccg
     2521 gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg gccgacagcg gcaaccgccg
     2581 gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt taaactctag aaccggtcat
     2641 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgttcga
     2701 actagatgct gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     2761 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     2821 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2881 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3421 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3721 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3781 cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga ggcaccgatc
     3841 tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt gtagatcact
     3901 acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt
     3961 tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga gcgaagaagt
     4021 ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga tgctagagta
     4081 agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg catcgtggta
     4141 tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc aagccgggtc
     4201 acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc gatcgttgtc
     4261 agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca caattctctt
     4321 accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac caagtcgttt
     4381 tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg ggacaacacc
     4441 gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc ggggcggaaa
     4501 gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct tgcacccagt
     4561 tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa
     4621 aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt
     4681 tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg ataagtaagt
     4741 aatattaagg tacgggaggt attggacagg ccgcaataaa atatctttat tttcattaca
     4801 tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa
     4861 aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac
     4921 atttctct
//
