LOCUS       sequence-193539-        6243 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 6243)
  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 6243)
  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 4
FEATURES             Location/Qualifiers
     source          1..6243
                     /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"
     polyA_signal    2409..2633
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2731..3319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2811..2830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3519..4310,4311..4379))
                     /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(4428..4663,4664..4863)
                     /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    4891..5115
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    5142..5366
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    6090..6138
                     /note="synthetic polyadenylation signal"
     misc_feature    6152..6243
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6192..6211
                     /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 acatgttcga ggcgactcta gaggatcgag ccccagctgg ttctttccgc
     2401 ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
     2461 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
     2521 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
     2581 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc
     2641 tgatcagcga gctctagaga attgatccca catgtgagca aaaggccagc aaaaggccag
     2701 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2761 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2821 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2881 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2941 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3001 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3061 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3121 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3181 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3241 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3301 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3361 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3421 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3481 gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt accagtgctt gatcagtgag
     3541 gcaccgatct cagcgatctg cctatttcgt tcgtccatag tggcctgact ccccgtcgtg
     3601 tagatcacta cgattcgtga gggcttacca tcaggcccca gcgcagcaat gatgccgcga
     3661 gagccgcgtt caccggcccc cgatttgtca gcaatgaacc agccagcagg gagggccgag
     3721 cgaagaagtg gtcctgctac tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat
     3781 gctagagtaa gaagttcgcc agtgagtagt ttccgaagag ttgtggccat tgctactggc
     3841 atcgtggtat cacgctcgtc gttcggtatg gcttcgttca actctggttc ccagcggtca
     3901 agccgggtca catgatcacc catattatga agaaatgcag tcagctcctt agggcctccg
     3961 atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc agcactacac
     4021 aattctctta ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga gtactcaacc
     4081 aagtcgtttt gtgagtagtg tatacggcga ccaagctgct cttgcccggc gtctatacgg
     4141 gacaacaccg cgccacatag cagtactttg aaagtgctca tcatcgggaa tcgttcttcg
     4201 gggcggaaag actcaaggat cttgccgcta ttgagatcca gttcgatata gcccactctt
     4261 gcacccagtt gatcttcagc atcttttact ttcaccagcg tttcggggtg tgcaaaaaca
     4321 ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg gatgctcata
     4381 ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt actagttctg ctgtatctct
     4441 aagatggctg gcacgtaaag agggtctcaa taaatatgcc ttgaattaat gaaaacagtg
     4501 gtttctgaat catggcctcc aaagagcatt tgccttttcc cctctcaccc ttctgaaggt
     4561 gcccaggctg ccctggcagg atgcctcttt ctctgtgggg tggcattctc tgctctctct
     4621 ctaatactct ttctttttct tccctcctct cccccaactg caggatgcct ttgtggaact
     4681 gtaaggcccc agcatgcggc ctctgtttga tttctcctgg ctgtctctga agactctgct
     4741 cagtttggcc ctggtgggag cttgcatcac cctgggtgcc tatctgggcc acaagtgaag
     4801 tcaacatgcc tgccccaaac aaatatgcaa aaggttcact aaagcagtag aaataatatg
     4861 catagcgcca gcacagtggt cgagcctcga ctgtgccttc tagttgccag ccatctgttg
     4921 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4981 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     5041 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     5101 cggtgggctc tatggcttct gaggcggaaa gctagactcg actgtgcctt ctagttgcca
     5161 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5221 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     5281 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     5341 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agcttcagcg atggcttcta
     5401 gctgcccggg tggcatccct gtgacccctc cccagtgcct cttctggccc tggaagttgc
     5461 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     5521 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     5581 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     5641 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagccttc
     5701 tagcttcagc gatggcttct agctgcccgg gtggcatccc tgtgacccct ccccagtgcc
     5761 tcttctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     5821 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     5881 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     5941 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     6001 gattctcctg cctcagcctc ccgagtacta gtacgtctct caaggataag taagtaatat
     6061 taaggtacgg gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt
     6121 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
     6181 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
     6241 tct
//
