LOCUS       sequence-121007-        4998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF9 Op x6 -- GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4998)
  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 4998)
  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-192
FEATURES             Location/Qualifiers
     source          1..4998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        126..157
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      184..193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(190..192,193..195,196..909)
                     /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(190..192,193..195,196..906)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(235..256)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(496..515)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     843..864
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    953..1074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(964..983)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1018..1037
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1268..1625
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1268..1459
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1291..1311)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1476..1611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1538..1557
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1656..2693
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2672..2680
                     /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    2717..2765
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2921..2938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3092..3680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3172..3191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3880..4671,4672..4740))
                     /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    4845..4893
                     /note="synthetic polyadenylation signal"
     misc_feature    4907..4998
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4947..4966
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gctgtagatg gagaaatgta gatggagaaa
       61 tgtagatgga gaaatgtaga tggagaaatg tagatggaga aatgtagatg gagaagctta
      121 gacactagag ggtatataat ggaagctcga cttccagctt ggcaatccgg tactgttggt
      181 aaagccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      241 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      301 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      361 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
      421 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      481 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
      541 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
      601 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
      661 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
      721 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
      781 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
      841 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
      901 tacaagtaat aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
      961 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1021 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1081 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     1141 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc
     1201 ttccgctgat ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     1261 ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     1321 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     1501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     1561 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     1621 gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta
     1681 ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg atgcagttgt
     1741 cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc tatgtactgc
     1801 gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca
     1861 gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag agcctgacat
     1921 actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa acagagctgc
     1981 ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc gacctcagtc
     2041 aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca acctggcggg
     2101 atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg atggacgaca
     2161 ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc gaagactgtc
     2221 ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg accgacaacg
     2281 gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt cagtacgagg
     2341 tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag actcgctact
     2401 tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac atgctgcgca
     2461 tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat gctgcctggg
     2521 ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt cgcacacaaa
     2581 tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg gccgacagcg
     2641 gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt taaactctag
     2701 aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     2761 gtgtgttcga actagatgct gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     2821 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     2881 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     2941 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3001 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3061 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3121 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3181 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3241 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3301 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3361 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3421 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3481 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3541 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3601 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3661 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3721 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3781 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3841 ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga
     3901 ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt
     3961 gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg
     4021 agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga
     4081 gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga
     4141 tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg
     4201 catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc
     4261 aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc
     4321 gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca
     4381 caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac
     4441 caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg
     4501 ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc
     4561 ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct
     4621 tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac
     4681 aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat
     4741 actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg
     4801 ataagtaagt aatattaagg tacgggaggt attggacagg ccgcaataaa atatctttat
     4861 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc
     4921 atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag
     4981 gtgccagaac atttctct
//
