LOCUS       sequence-183860-        5924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector to measure enhancer activity of candidates from a DNA library
            by STARR-seq..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5924)
  AUTHORS   Muerdter F, Boryn LM, Woodfin AR, Neumayr C, Rath M, Zabidi MA, 
            Pagani M, Haberle V, Kazmar T, Catarino RR, Schernhuber K, Arnold 
            CD, Stark A
  TITLE     Resolving systematic errors in widely used enhancer activity assays 
            in human cells.
  JOURNAL   Nat Methods. 2017 Dec 11. pii: nmeth.4534. doi: 10.1038/nmeth.4534.
  PUBMED    29256496
REFERENCE   2  (bases 1 to 5924)
  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. 2017 Dec 11. pii: nmeth.4534. doi: 10.1038/nmeth.4534."
COMMENT     Sequence Label: hSTARR-seq_SCP1 vector_blocking 3
FEATURES             Location/Qualifiers
     source          1..5924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          140..272
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     255..274
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        join(407..412,413..430,431..437)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             491..1150
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(557..576)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1492..1797
                     /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     1676..1695
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1875..1996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1886..1905)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1940..1959
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2241..2258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2412..3000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2492..2511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3200..3991,3992..4060))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    join(4109..4344,4345..4544)
                     /label=Bcl-2 SA
                     /note="splice acceptor site from the intron 2/exon 3 
                     junction of the human Bcl-2 gene (Seto et al., 1988)"
     polyA_signal    4572..4796
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4823..5047
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    5771..5819
                     /note="synthetic polyadenylation signal"
     misc_feature    5833..5924
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5873..5892
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcccta gggtacttat ataagggggt gggggcgcgt
       61 tcgtcctcag tcgcgatcga acactcgagc cgagcagacg tgcctacgga ccggggcccg
      121 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
      181 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
      241 tactgacatc cactttgcct ttctctccac aggtgtccac cctgcaggct taagcatggc
      301 tagcaaagga gaagaactct tcactggagt tgtcccaatt cttgttgaat tagattgatc
      361 tagagcatgc accggtgata tcgcggccgc attaggcacc ccaggcttta cactttatgc
      421 ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt caggagctaa
      481 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
      541 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
      601 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
      661 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat
      721 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
      781 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
      841 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
      901 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
      961 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     1021 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1081 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     1141 cggggcgtaa acgcgtggat ccggcttact aaaagccaga taacagtatg cgtatttgcg
     1201 cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag
     1261 aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc
     1321 aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc
     1381 gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc
     1441 ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt
     1501 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
     1561 attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
     1621 gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
     1681 atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc
     1741 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg
     1801 tcaggctccc ttatacacag ccagtctgca ggatatcgtc gacgaattcg gccggccgct
     1861 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1921 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1981 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2041 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     2101 tccgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     2161 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     2221 cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2281 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2341 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2401 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2461 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2521 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2581 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2641 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2701 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2761 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2821 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2881 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2941 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3001 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3061 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag cggccgcaaa
     3181 tgctaaacca ctgcagtggt taccagtgct tgatcagtga ggcaccgatc tcagcgatct
     3241 gcctatttcg ttcgtccata gtggcctgac tccccgtcgt gtagatcact acgattcgtg
     3301 agggcttacc atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt tcaccggccc
     3361 ccgatttgtc agcaatgaac cagccagcag ggagggccga gcgaagaagt ggtcctgcta
     3421 ctttgtccgc ctccatccag tctatgagct gctgtcgtga tgctagagta agaagttcgc
     3481 cagtgagtag tttccgaaga gttgtggcca ttgctactgg catcgtggta tcacgctcgt
     3541 cgttcggtat ggcttcgttc aactctggtt cccagcggtc aagccgggtc acatgatcac
     3601 ccatattatg aagaaatgca gtcagctcct tagggcctcc gatcgttgtc agaagtaagt
     3661 tggccgcggt gttgtcgctc atggtaatgg cagcactaca caattctctt accgtcatgc
     3721 catccgtaag atgcttttcc gtgaccggcg agtactcaac caagtcgttt tgtgagtagt
     3781 gtatacggcg accaagctgc tcttgcccgg cgtctatacg ggacaacacc gcgccacata
     3841 gcagtacttt gaaagtgctc atcatcggga atcgttcttc ggggcggaaa gactcaagga
     3901 tcttgccgct attgagatcc agttcgatat agcccactct tgcacccagt tgatcttcag
     3961 catcttttac tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa aatgccgcaa
     4021 agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt tttcaatatt
     4081 attgaagcat ttatcagggt tactagttct gctgtatctc taagatggct ggcacgtaaa
     4141 gagggtctca ataaatatgc cttgaattaa tgaaaacagt ggtttctgaa tcatggcctc
     4201 caaagagcat ttgccttttc ccctctcacc cttctgaagg tgcccaggct gccctggcag
     4261 gatgcctctt tctctgtggg gtggcattct ctgctctctc tctaatactc tttctttttc
     4321 ttccctcctc tcccccaact gcaggatgcc tttgtggaac tgtaaggccc cagcatgcgg
     4381 cctctgtttg atttctcctg gctgtctctg aagactctgc tcagtttggc cctggtggga
     4441 gcttgcatca ccctgggtgc ctatctgggc cacaagtgaa gtcaacatgc ctgccccaaa
     4501 caaatatgca aaaggttcac taaagcagta gaaataatat gcatagcgcc agcacagtgg
     4561 tcgagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4621 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4681 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4741 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4801 tgaggcggaa agctagactc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4861 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4921 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4981 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     5041 tctatggctt ctgaggcgga aagcttcagc gatggcttct agctgcccgg gtggcatccc
     5101 tgtgacccct ccccagtgcc tcttctggcc ctggaagttg ccactccagt gcccaccagc
     5161 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     5221 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     5281 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     5341 tccgcctcct gggttcaagc gattctcctg cctcagcctt ctagcttcag cgatggcttc
     5401 tagctgcccg ggtggcatcc ctgtgacccc tccccagtgc ctcttctggc cctggaagtt
     5461 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     5521 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     5581 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     5641 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     5701 cccgagtact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg ggaggtattg
     5761 gacaggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     5821 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa
     5881 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctct
//
