LOCUS       sequence-125112-        6127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF9 Op x6 -- GFP-IRES-NTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6127)
  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 6127)
  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-352
FEATURES             Location/Qualifiers
     source          1..6127
                     /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    2082..2203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2093..2112)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2147..2166
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2397..2754
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2397..2588
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2420..2440)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2605..2740
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2667..2686
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2785..3822
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3801..3809
                     /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    3846..3894
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4050..4067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4221..4809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4301..4320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5009..5800,5801..5869))
                     /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    5974..6022
                     /note="synthetic polyadenylation signal"
     misc_feature    6036..6127
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6076..6095
                     /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 tacaagtaaa gcaggtttcc ccaatgacac aaaacgtgca acttgaaact ccgcctggtc
      961 tttccaggtc tagaggggta acactttgta ctgcgtttgg ctccacgctc gatccactgg
     1021 cgagtgttag taacagcact gttgcttcgt agcggagcat gacggccgtg ggaactcctc
     1081 cttggtaaca aggacccacg gggccaaaag ccacgcccac acgggcccgt catgtgtgca
     1141 accccagcac ggcgacttta ctgcgaaacc cactttaaag tgacattgaa actggtaccc
     1201 acacactggt gacaggctaa ggatgccctt caggtacccc gaggtaacac gcgacactcg
     1261 ggatctgaga aggggactgg ggcttctata aaagcgctcg gtttaaaaag cttctatgcc
     1321 tgaataggtg accggaggtc ggcacctttc cgttgcaatt actgacccta tgaatacact
     1381 cgagatggac atcatcagcg tggctctgaa gaggcactcc accaaggctt tcgacgcttc
     1441 caagaaactg acccctgaac aggccgagca gatcaagacc ctgctccagt acagccctag
     1501 ctccaccaac agccagcctt ggcacttcat cgtggctagc accgaggaag gcaaagctag
     1561 ggtggctaag agcgccgctg gcaactacgt gttcaacgag aggaagatgc tggatgctag
     1621 ccacgtggtg gtgttctgcg ctaagaccgc catggacgat gtgtggctga agctggtggt
     1681 ggatcaggaa gatgctgatg gcaggttcgc tacccctgaa gctaaggccg ctaacgacaa
     1741 gggcaggaag ttcttcgccg acatgcacag gaaggatctg cacgatgatg ctgagtggat
     1801 ggccaagcag gtgtacctga acgtgggcaa cttcctgctc ggcgtggctg ccctgggcct
     1861 cgatgctgtg cccatcgaag gcttcgatgc tgctatcctg gatgccgagt tcggcctgaa
     1921 ggagaaaggc tacaccagcc tggtggtggt gcctgtgggc caccacagcg tggaggactt
     1981 caacgctacc ctgcctaaga gcaggctgcc ccagaacatc accctgaccg aggtgtgata
     2041 attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2101 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2161 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2221 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     2281 ctacaaatgt ggtaaaatcg ataaggatcc gtttgcgtat tgggcgctct tccgctgatc
     2341 tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagc taccttctga
     2401 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2461 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2521 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2581 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2641 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     2701 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     2761 gattcttctg acactagcgc caccatgaag aagcccgaac tcaccgctac cagcgttgaa
     2821 aaatttctca tcgagaagtt cgacagtgtg agcgacctga tgcagttgtc ggagggcgaa
     2881 gagagccgag ccttcagctt cgatgtcggc ggacgcggct atgtactgcg ggtgaatagc
     2941 tgcgctgatg gcttctacaa agaccgctac gtgtaccgcc acttcgccag cgctgcacta
     3001 cccatccccg aagtgttgga catcggcgag ttcagcgaga gcctgacata ctgcatcagt
     3061 agacgcgccc aaggcgttac tctccaagac ctccccgaaa cagagctgcc tgctgtgtta
     3121 cagcctgtcg ccgaagctat ggatgctatt gccgccgccg acctcagtca aaccagcggc
     3181 ttcggcccat tcgggcccca aggcatcggc cagtacacaa cctggcggga tttcatttgc
     3241 gccattgctg atccccatgt ctaccactgg cagaccgtga tggacgacac cgtgtccgcc
     3301 agcgtagctc aagccctgga cgaactgatg ctgtgggccg aagactgtcc cgaggtgcgc
     3361 cacctcgtcc atgccgactt cggcagcaac aacgtcctga ccgacaacgg ccgcatcacc
     3421 gccgtaatcg actggtccga agctatgttc ggggacagtc agtacgaggt ggccaacatc
     3481 ttcttctggc ggccctggct ggcttgcatg gagcagcaga ctcgctactt cgagcgccgg
     3541 catcccgagc tggccggcag ccctcgtctg cgagcctaca tgctgcgcat cggcctggat
     3601 cagctctacc agagcctcgt ggacggcaac ttcgacgatg ctgcctgggc tcaaggccgc
     3661 tgcgatgcca tcgtccgcag cggggccggc accgtcggtc gcacacaaat cgctcgccgg
     3721 agcgcagccg tatggaccga cggctgcgtc gaggtgctgg ccgacagcgg caaccgccgg
     3781 cccagtacac gaccgcgcgc taaggaggta ggtcgagttt aaactctaga accggtcatg
     3841 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgttcgaa
     3901 ctagatgctg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     3961 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     4021 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4081 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4141 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4201 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4261 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4321 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4381 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4441 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4501 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4561 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4621 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4681 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4741 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4801 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4861 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4921 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc
     4981 ggccgcaaat gctaaaccac tgcagtggtt accagtgctt gatcagtgag gcaccgatct
     5041 cagcgatctg cctatttcgt tcgtccatag tggcctgact ccccgtcgtg tagatcacta
     5101 cgattcgtga gggcttacca tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt
     5161 caccggcccc cgatttgtca gcaatgaacc agccagcagg gagggccgag cgaagaagtg
     5221 gtcctgctac tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat gctagagtaa
     5281 gaagttcgcc agtgagtagt ttccgaagag ttgtggccat tgctactggc atcgtggtat
     5341 cacgctcgtc gttcggtatg gcttcgttca actctggttc ccagcggtca agccgggtca
     5401 catgatcacc catattatga agaaatgcag tcagctcctt agggcctccg atcgttgtca
     5461 gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc agcactacac aattctctta
     5521 ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga gtactcaacc aagtcgtttt
     5581 gtgagtagtg tatacggcga ccaagctgct cttgcccggc gtctatacgg gacaacaccg
     5641 cgccacatag cagtactttg aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag
     5701 actcaaggat cttgccgcta ttgagatcca gttcgatata gcccactctt gcacccagtt
     5761 gatcttcagc atcttttact ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa
     5821 atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg gatgctcata ctcgtccttt
     5881 ttcaatatta ttgaagcatt tatcagggtt actagtacgt ctctcaagga taagtaagta
     5941 atattaaggt acgggaggta ttggacaggc cgcaataaaa tatctttatt ttcattacat
     6001 ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa
     6061 acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca
     6121 tttctct
//
