LOCUS       sequence-129009-        6692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  flySTARR-seq validation vector with Gateway cassette downstream of 
            the firefly luciferase gene and the DSCP core promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  AUTHORS   Zabidi MA, Arnold CD, Schernhuber K, Pagani M, Rath M, Frank O, 
            Stark A
  TITLE     Enhancer-core-promoter specificity separates developmental and 
            housekeeping gene regulation.
  JOURNAL   Nature. 2015 Feb 26;518(7540):556-9. doi: 10.1038/nature13994. Epub 
            2014 Dec 15.
  PUBMED    25517091
REFERENCE   2  (bases 1 to 6692)
  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"; doi: 
            "10.1038/nature13994"; journalName: "Nature"; date: "2015-02-26- 
            26"; volume: "518"; issue: "7540"; pages: "556-9"
COMMENT     Sequence Label: pGL3-DSCP-Gateway
FEATURES             Location/Qualifiers
     source          1..6692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    7..55
                     /note="synthetic polyadenylation signal"
     misc_feature    69..160
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     109..128
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      418..427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             424..2076
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(490..508)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2117..2238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2128..2147)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2182..2201
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    2473..2597
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2622..2627,2628..2645,2646..2652)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2706..3365
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2772..2791)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3707..4012
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3891..3910
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4053..4177)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4191..4208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4362..4950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4442..4461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5121..5912,5913..5981))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5744..5763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5982..6086)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6113..6568
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6200..6219)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6410..6431
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
       61 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
      121 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt accgagctct
      181 tacgcgtcca tgcatgctat gggaattccc atcagacgtc tggctcgaga tctgcgatcg
      241 agctcgcccg gggatcgagc gcagcggtat aaaagggcgc ggggtggctg agagcatcag
      301 ttgtgaatga atgttcgagc cgagcagacg tgccgctgcc ttcgttaata tcctttgaat
      361 aagccaactt tgaatcacaa gacgcatacc aaacggcatt ccggtactgt tggtaaagcc
      421 accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat
      481 ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca
      541 attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg
      601 tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc
      661 gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga
      721 gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc
      781 atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg
      841 caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag
      901 ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac
      961 gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct
     1021 ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc
     1081 tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt
     1141 gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga
     1201 tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat
     1261 tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact
     1321 ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc
     1381 tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga
     1441 tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg
     1501 ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg
     1561 aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc
     1621 ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat
     1681 tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag
     1741 tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc
     1801 caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa
     1861 cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg
     1921 gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg
     1981 gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc
     2041 ataaaggcca agaagggcgg aaagatcgcc gtgtaattct agagtcgggg cggccggccg
     2101 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2161 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2221 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2281 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     2341 gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     2401 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     2461 ggtgccggca gcacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     2521 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     2581 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     2641 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
     2701 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     2761 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     2821 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     2881 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     2941 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     3001 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     3061 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     3121 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     3181 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     3241 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     3301 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     3361 cgtaaacgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     3421 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta
     3481 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     3541 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     3601 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     3661 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     3721 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     3781 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     3841 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     3901 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     3961 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     4021 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     4081 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     4141 tttacgtttc tcgttcagct ttcttgtaca aagtggtgct cttccgcttc ctcgctcact
     4201 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4261 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4321 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4381 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4441 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4501 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4561 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4621 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4681 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4741 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4801 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4861 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4921 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4981 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5041 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5101 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5161 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5221 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5281 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5341 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5401 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5461 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5521 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5581 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5641 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5701 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5761 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5821 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5881 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5941 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6001 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6061 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     6121 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6181 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6241 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6301 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6361 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6421 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6481 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6541 tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg
     6601 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta
     6661 agtaatatta aggtacggga ggtacttgga gc
//
