LOCUS       sequence-121004-        4939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF7-ZF8 Op 2gap -- GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4939)
  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 4939)
  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-165
FEATURES             Location/Qualifiers
     source          1..4939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        67..98
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      125..134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(131..133,134..136,137..850)
                     /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(131..133,134..136,137..847)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(176..197)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(437..456)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     784..805
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    894..1015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(905..924)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     959..978
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1209..1566
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1209..1400
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1232..1252)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1417..1552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1479..1498
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1597..2634
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2613..2621
                     /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    2658..2706
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2862..2879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3033..3621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3113..3132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3821..4612,4613..4681))
                     /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    4786..4834
                     /note="synthetic polyadenylation signal"
     misc_feature    4848..4939
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4888..4907
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcagctggag ggtcgaagtg gtccaagctt
       61 agacactaga gggtatataa tggaagctcg acttccagct tggcaatccg gtactgttgg
      121 taaagccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      181 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      241 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      301 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      361 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      421 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      481 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      541 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
      601 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
      661 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
      721 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
      781 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
      841 gtacaagtaa taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata
      901 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      961 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1021 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1081 caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtttgcgt attgggcgct
     1141 cttccgctga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     1201 gctaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     1261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1321 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1381 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     1441 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     1501 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     1561 tgcaaaaagc tcgattcttc tgacactagc gccaccatga agaagcccga actcaccgct
     1621 accagcgttg aaaaatttct catcgagaag ttcgacagtg tgagcgacct gatgcagttg
     1681 tcggagggcg aagagagccg agccttcagc ttcgatgtcg gcggacgcgg ctatgtactg
     1741 cgggtgaata gctgcgctga tggcttctac aaagaccgct acgtgtaccg ccacttcgcc
     1801 agcgctgcac tacccatccc cgaagtgttg gacatcggcg agttcagcga gagcctgaca
     1861 tactgcatca gtagacgcgc ccaaggcgtt actctccaag acctccccga aacagagctg
     1921 cctgctgtgt tacagcctgt cgccgaagct atggatgcta ttgccgccgc cgacctcagt
     1981 caaaccagcg gcttcggccc attcgggccc caaggcatcg gccagtacac aacctggcgg
     2041 gatttcattt gcgccattgc tgatccccat gtctaccact ggcagaccgt gatggacgac
     2101 accgtgtccg ccagcgtagc tcaagccctg gacgaactga tgctgtgggc cgaagactgt
     2161 cccgaggtgc gccacctcgt ccatgccgac ttcggcagca acaacgtcct gaccgacaac
     2221 ggccgcatca ccgccgtaat cgactggtcc gaagctatgt tcggggacag tcagtacgag
     2281 gtggccaaca tcttcttctg gcggccctgg ctggcttgca tggagcagca gactcgctac
     2341 ttcgagcgcc ggcatcccga gctggccggc agccctcgtc tgcgagccta catgctgcgc
     2401 atcggcctgg atcagctcta ccagagcctc gtggacggca acttcgacga tgctgcctgg
     2461 gctcaaggcc gctgcgatgc catcgtccgc agcggggccg gcaccgtcgg tcgcacacaa
     2521 atcgctcgcc ggagcgcagc cgtatggacc gacggctgcg tcgaggtgct ggccgacagc
     2581 ggcaaccgcc ggcccagtac acgaccgcgc gctaaggagg taggtcgagt ttaaactcta
     2641 gaaccggtca tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     2701 tgtgtgttcg aactagatgc tgtcgaccga tgcccttgag agccttcaac ccagtcagct
     2761 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     2821 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     2881 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2941 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3001 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3061 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3121 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3181 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3241 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3301 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3361 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3421 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3481 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3541 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3601 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3661 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3781 tggtctgaca gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc ttgatcagtg
     3841 aggcaccgat ctcagcgatc tgcctatttc gttcgtccat agtggcctga ctccccgtcg
     3901 tgtagatcac tacgattcgt gagggcttac catcaggccc cagcgcagca atgatgccgc
     3961 gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa ccagccagca gggagggccg
     4021 agcgaagaag tggtcctgct actttgtccg cctccatcca gtctatgagc tgctgtcgtg
     4081 atgctagagt aagaagttcg ccagtgagta gtttccgaag agttgtggcc attgctactg
     4141 gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt caactctggt tcccagcggt
     4201 caagccgggt cacatgatca cccatattat gaagaaatgc agtcagctcc ttagggcctc
     4261 cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct catggtaatg gcagcactac
     4321 acaattctct taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc gagtactcaa
     4381 ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg gcgtctatac
     4441 gggacaacac cgcgccacat agcagtactt tgaaagtgct catcatcggg aatcgttctt
     4501 cggggcggaa agactcaagg atcttgccgc tattgagatc cagttcgata tagcccactc
     4561 ttgcacccag ttgatcttca gcatctttta ctttcaccag cgtttcgggg tgtgcaaaaa
     4621 caggcaagca aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt tggatgctca
     4681 tactcgtcct ttttcaatat tattgaagca tttatcaggg ttactagtac gtctctcaag
     4741 gataagtaag taatattaag gtacgggagg tattggacag gccgcaataa aatatcttta
     4801 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
     4861 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
     4921 ggtgccagaa catttctct
//
