LOCUS       sequence-129008-        6638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  flySTARR-seq validation vector with Gateway cassette upstream 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 6638)
  AUTHORS   Arnold CD, Gerlach D, Stelzer C, Boryn LM, Rath M, Stark A
  TITLE     Genome-wide quantitative enhancer activity maps identified by 
            STARR-seq.
  JOURNAL   Science. 2013 Mar 1;339(6123):1074-7. doi: 10.1126/science.1232542. 
            Epub 2013 Jan 17.
  PUBMED    23328393
REFERENCE   2  (bases 1 to 6638)
  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: 
            "10.1126/science.1232542"; journalName: "Science"; date: "2013-03-1-
            1"; volume: "339"; issue: "6123"; pages: "1074-7"
COMMENT     Sequence Label: pGL3-Gateway-DSCP
FEATURES             Location/Qualifiers
     source          1..6638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      993..1448
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1080..1099)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1290..1311
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1579..1627
                     /note="synthetic polyadenylation signal"
     misc_feature    1641..1732
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1681..1700
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    1746..1870
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1895..1900,1901..1918,1919..1925)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1979..2638
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2045..2064)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2980..3285
                     /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     3164..3183
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3326..3450)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3641..3650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3647..5299
                     /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(3713..3731)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5340..5461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5351..5370)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5405..5424
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5709..5726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5880..6468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5960..5979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     1021 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1081 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1141 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1201 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1261 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1321 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     1381 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     1441 tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     1501 ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga
     1561 ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     1621 ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa
     1681 ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgatag
     1741 gtaccacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata aatatcaata
     1801 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
     1861 agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     1921 taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg aagctaaaat
     1981 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     2041 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     2101 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     2161 cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga
     2221 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     2281 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     2341 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     2401 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     2461 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     2521 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     2581 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaaac
     2641 gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg
     2701 cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag gtatgctatg
     2761 aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg
     2821 atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg
     2881 ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa
     2941 tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc
     3001 tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg
     3061 cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc
     3121 cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat
     3181 atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa
     3241 aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc aggctccctt
     3301 atacacagcc agtctgcagg tcgaccatag tgactggata tgttgtgttt tacagtatta
     3361 tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat cattttacgt
     3421 ttctcgttca gctttcttgt acaaagtggt agatctgcga tcgagctcgc ccggggatcg
     3481 agcgcagcgg tataaaaggg cgcggggtgg ctgagagcat cagttgtgaa tgaatgttcg
     3541 agccgagcag acgtgccgct gccttcgtta atatcctttg aataagccaa ctttgaatca
     3601 caagacgcat accaaacggc attccggtac tgttggtaaa gccaccatgg aagacgccaa
     3661 aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca
     3721 actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc
     3781 acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga
     3841 agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc
     3901 tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc
     3961 gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt
     4021 ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat
     4081 catccaaaaa attattatca tggattctaa aacggattac cagggatttc agtcgatgta
     4141 cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc
     4201 cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc
     4261 taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc
     4321 tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca
     4381 cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat
     4441 gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc
     4501 gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga
     4561 tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga
     4621 agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac
     4681 tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt
     4741 tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa
     4801 tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc
     4861 ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag acatagctta
     4921 ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa
     4981 aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt
     5041 cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt
     5101 tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca
     5161 agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg
     5221 tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg
     5281 cggaaagatc gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     5341 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     5401 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5461 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     5521 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc
     5581 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     5641 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     5701 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5761 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5821 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5881 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5941 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6001 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6061 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6121 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6181 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6241 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6301 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6361 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6421 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6481 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6541 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6601 tcaatctaaa gtatatatga gtaaacttgg tctgacag
//
