LOCUS       sequence-129002-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  flySTARR-seq screening vector with hsp70 core promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  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 5570)
  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: pSTARR-seq_fly-hsp70
FEATURES             Location/Qualifiers
     source          1..5570
                     /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      365..374
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          380..446
                     /label=Mhc intron
                     /note="modified Drosophila myosin heavy chain intron 16 
                     (Pfeiffer et al., 2010)"
     primer_bind     complement(494..522)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1117..1137
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        join(1229..1234,1235..1252,1253..1259)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1313..1972
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1379..1398)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2314..2619
                     /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     2498..2517
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    2697..2818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2708..2727)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2762..2781
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3063..3080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3234..3822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3314..3333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3993..4784,4785..4853))
                     /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     4616..4635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4854..4958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4985..5440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5072..5091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5282..5303
                     /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 ctgagcgccg gagtataaat agaggcgctt cgtctacgga
      241 gcgacaattc aattcaaaca agcaaagtga acacgtcgct aagcgaaagc taagcaaata
      301 aacaagcgca gctgaacaag ctaaacaatc tgcagaagct tggcattccg gtactgttgg
      361 taaagccacc atggaaaagg taggttcaac cactgatgcc taggcacacc gaaacgacta
      421 accctaattc ttatccttta cttcaggcca aagcttaagc atggctagca aaggagaaga
      481 actcttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta acggccacaa
      541 gttctctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta ccctgaagtt
      601 catctgcact actggcaaac tgcctgttcc ctggccaaca ctagtcacta ctctgtgcta
      661 tggtgttcaa tgcttttcaa gatacccgga tcatatgaaa cggcatgact ttttcaagag
      721 tgccatgccc gaaggttatg tacaggaaag gaccatcttc ttcaaagatg acggcaacta
      781 caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa
      841 aggtattgac ttcaaggaag atggcaacat tctgggacac aaattggaat acaactataa
      901 ctcacacaat gtatacatca tggcagacaa acaaaagaat ggaatcaaag cgaacttcaa
      961 gacccgccac aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac
     1021 tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc
     1081 cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc
     1141 tgctgggatt acacatggca tggatgaact gtacaactga tctagagcat gcaccggtga
     1201 tatcgcggcc gcattaggca ccccaggctt tacactttat gcttccggct cgtataatgt
     1261 gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta aaatggagaa
     1321 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     1381 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     1441 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     1501 tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg gtgagctggt
     1561 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     1621 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     1681 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     1741 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     1801 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
     1861 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     1921 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aaacgcgtgg
     1981 atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt tttgcggtat
     2041 aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc tatgaagcag
     2101 cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata tatgatgtca
     2161 atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac
     2221 gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg
     2281 gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta cacctataaa
     2341 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgcccggg
     2401 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa
     2461 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc
     2521 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac
     2581 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc ccttatacac
     2641 agccagtctg caggatatcg tcgacgaatt cggccggccg cttcgagcag acatgataag
     2701 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2761 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     2821 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     2881 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgaccg atgcccttga
     2941 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     3001 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct
     3061 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4981 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5401 tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc attcaggctg
     5461 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcccaagcta
     5521 ccatgataag taagtaatat taaggtacgg gaggtacttg gagcggccgc
//
