LOCUS       sequence-136739-        5543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  flySTARR-seq screening vector with x16 core promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5543)
  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 5543)
  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: pSTARR-seq_fly-x16
FEATURES             Location/Qualifiers
     source          1..5543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      341..350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          356..422
                     /label=Mhc intron
                     /note="modified Drosophila myosin heavy chain intron 16 
                     (Pfeiffer et al., 2010)"
     primer_bind     complement(467..495)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1090..1110
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        join(1202..1207,1208..1225,1226..1232)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1286..1945
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1352..1371)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2287..2592
                     /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     2471..2490
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    2670..2791
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2681..2700)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2735..2754
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3036..3053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3207..3795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3287..3306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3966..4757,4758..4826))
                     /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     4589..4608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4827..4931)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4958..5413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5045..5064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5255..5276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcttacgcg
      181 tgctagcccg ggctcgagat ctgcgatcct tttgctgacc gttacccgcg gattgagcat
      241 aacttttggt atatttttag cgagtccgca gtttctttta atggtctgtc caacggtcac
      301 tctgtttgcc ctgcaggggc attccggtac tgttggtaaa gccaccatgg aaaaggtagg
      361 ttcaaccact gatgcctagg cacaccgaaa cgactaaccc taattcttat cctttacttc
      421 aggccactta agcatggcta gcaaaggaga agaactcttc actggagttg tcccaattct
      481 tgttgaatta gatggtgatg ttaacggcca caagttctct gtcagtggag agggtgaagg
      541 tgatgcaaca tacggaaaac ttaccctgaa gttcatctgc actactggca aactgcctgt
      601 tccctggcca acactagtca ctactctgtg ctatggtgtt caatgctttt caagataccc
      661 ggatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
      721 aaggaccatc ttcttcaaag atgacggcaa ctacaagaca cgtgctgaag tcaagtttga
      781 aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gacttcaagg aagatggcaa
      841 cattctggga cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga
      901 caaacaaaag aatggaatca aagcgaactt caagacccgc cacaacattg aagatggaag
      961 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     1021 accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag
     1081 agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga
     1141 actgtacaac tgatctagag catgcaccgg tgatatcgcg gccgcattag gcaccccagg
     1201 ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag
     1261 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
     1321 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     1381 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     1441 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     1501 attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     1561 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     1621 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     1681 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
     1741 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     1801 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     1861 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     1921 cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca
     1981 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
     2041 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
     2101 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
     2161 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
     2221 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
     2281 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     2341 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
     2401 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     2461 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     2521 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     2581 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggata tcgtcgacga
     2641 attcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2701 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2761 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2821 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2881 taaaatcgat aaggatccga ccgatgccct tgagagcctt caacccagtc agctccttcc
     2941 ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac
     3001 tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3061 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3121 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3181 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3661 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3841 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3901 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3961 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4021 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4081 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4141 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4201 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4261 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4321 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4381 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4441 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4501 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4561 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4621 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4681 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4741 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4801 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4861 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4921 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     4981 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5041 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5101 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5161 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5221 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5281 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5341 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5401 acgcttacaa tttgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5461 cgggcctctt cgctattacg ccagcccaag ctaccatgat aagtaagtaa tattaaggta
     5521 cgggaggtac ttggagcggc cgc
//
