LOCUS       sequence-120994-        4937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF10-ZF6 Op 0gap --GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4937)
  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 4937)
  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-160
FEATURES             Location/Qualifiers
     source          1..4937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        65..96
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      123..132
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(129..131,132..134,135..848)
                     /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(129..131,132..134,135..845)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(174..195)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(435..454)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     782..803
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    892..1013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(903..922)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     957..976
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1207..1564
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1207..1398
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1230..1250)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1415..1550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1477..1496
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1595..2632
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2611..2619
                     /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    2656..2704
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2860..2877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3031..3619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3111..3130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3819..4610,4611..4679))
                     /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    4784..4832
                     /note="synthetic polyadenylation signal"
     misc_feature    4846..4937
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4886..4905
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gctgaagaag ctgcaggagg tgaagcttag
       61 acactagagg gtatataatg gaagctcgac ttccagcttg gcaatccggt actgttggta
      121 aagccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      181 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      241 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      301 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      361 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      421 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      481 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      541 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      601 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      661 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      721 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      781 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      841 acaagtaata attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
      901 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
      961 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1021 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     1081 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtttgcgtat tgggcgctct
     1141 tccgctgatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagc
     1201 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     1261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     1321 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1381 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     1441 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     1501 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     1561 caaaaagctc gattcttctg acactagcgc caccatgaag aagcccgaac tcaccgctac
     1621 cagcgttgaa aaatttctca tcgagaagtt cgacagtgtg agcgacctga tgcagttgtc
     1681 ggagggcgaa gagagccgag ccttcagctt cgatgtcggc ggacgcggct atgtactgcg
     1741 ggtgaatagc tgcgctgatg gcttctacaa agaccgctac gtgtaccgcc acttcgccag
     1801 cgctgcacta cccatccccg aagtgttgga catcggcgag ttcagcgaga gcctgacata
     1861 ctgcatcagt agacgcgccc aaggcgttac tctccaagac ctccccgaaa cagagctgcc
     1921 tgctgtgtta cagcctgtcg ccgaagctat ggatgctatt gccgccgccg acctcagtca
     1981 aaccagcggc ttcggcccat tcgggcccca aggcatcggc cagtacacaa cctggcggga
     2041 tttcatttgc gccattgctg atccccatgt ctaccactgg cagaccgtga tggacgacac
     2101 cgtgtccgcc agcgtagctc aagccctgga cgaactgatg ctgtgggccg aagactgtcc
     2161 cgaggtgcgc cacctcgtcc atgccgactt cggcagcaac aacgtcctga ccgacaacgg
     2221 ccgcatcacc gccgtaatcg actggtccga agctatgttc ggggacagtc agtacgaggt
     2281 ggccaacatc ttcttctggc ggccctggct ggcttgcatg gagcagcaga ctcgctactt
     2341 cgagcgccgg catcccgagc tggccggcag ccctcgtctg cgagcctaca tgctgcgcat
     2401 cggcctggat cagctctacc agagcctcgt ggacggcaac ttcgacgatg ctgcctgggc
     2461 tcaaggccgc tgcgatgcca tcgtccgcag cggggccggc accgtcggtc gcacacaaat
     2521 cgctcgccgg agcgcagccg tatggaccga cggctgcgtc gaggtgctgg ccgacagcgg
     2581 caaccgccgg cccagtacac gaccgcgcgc taaggaggta ggtcgagttt aaactctaga
     2641 accggtcatg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     2701 tgtgttcgaa ctagatgctg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     2761 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     2821 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     2881 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2941 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3001 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3061 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3121 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3181 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3241 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3301 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3361 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3421 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3481 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3541 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3601 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3661 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3721 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3781 gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt accagtgctt gatcagtgag
     3841 gcaccgatct cagcgatctg cctatttcgt tcgtccatag tggcctgact ccccgtcgtg
     3901 tagatcacta cgattcgtga gggcttacca tcaggcccca gcgcagcaat gatgccgcga
     3961 gagccgcgtt caccggcccc cgatttgtca gcaatgaacc agccagcagg gagggccgag
     4021 cgaagaagtg gtcctgctac tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat
     4081 gctagagtaa gaagttcgcc agtgagtagt ttccgaagag ttgtggccat tgctactggc
     4141 atcgtggtat cacgctcgtc gttcggtatg gcttcgttca actctggttc ccagcggtca
     4201 agccgggtca catgatcacc catattatga agaaatgcag tcagctcctt agggcctccg
     4261 atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc agcactacac
     4321 aattctctta ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga gtactcaacc
     4381 aagtcgtttt gtgagtagtg tatacggcga ccaagctgct cttgcccggc gtctatacgg
     4441 gacaacaccg cgccacatag cagtactttg aaagtgctca tcatcgggaa tcgttcttcg
     4501 gggcggaaag actcaaggat cttgccgcta ttgagatcca gttcgatata gcccactctt
     4561 gcacccagtt gatcttcagc atcttttact ttcaccagcg tttcggggtg tgcaaaaaca
     4621 ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg gatgctcata
     4681 ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt actagtacgt ctctcaagga
     4741 taagtaagta atattaaggt acgggaggta ttggacaggc cgcaataaaa tatctttatt
     4801 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca
     4861 tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg
     4921 tgccagaaca tttctct
//
