LOCUS       sequence-193172-        4318 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 4318)
  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 4318)
  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
FEATURES             Location/Qualifiers
     source          1..4318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          181..313
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     296..315
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        join(448..453,454..471,472..478)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             532..1191
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(598..617)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1533..1838
                     /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     1717..1736
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1916..2037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1927..1946)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1981..2000
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2282..2299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2453..3041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2533..2552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3241..4032,4033..4101))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4206..4254
                     /note="synthetic polyadenylation signal"
     misc_feature    join(4268..4318,1..41)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     join(4308..4318,1..9)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggctgtcccc agtgcaagtg caggtgccag aacatttctc tggcctaact ggccggtacc
       61 tgagctccct agggtactta tataaggggg tgggggcgcg ttcgtcctca gtcgcgatcg
      121 aacactcgag ccgagcagac gtgcctacgg accggggccc ggtcgtgagg cactgggcag
      181 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
      241 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
      301 tttctctcca caggtgtcca ccctgcaggc ttaagcatgg ctagcaaagg agaagaactc
      361 ttcactggag ttgtcccaat tcttgttgaa ttagattgat ctagagcatg caccggtgat
      421 atcgcggccg cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg
      481 tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa
      541 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
      601 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
      661 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
      721 gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg
      781 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
      841 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
      901 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
      961 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1021 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     1081 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     1141 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta aacgcgtgga
     1201 tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata
     1261 agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc
     1321 gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa
     1381 tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg
     1441 ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg
     1501 ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa
     1561 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc
     1621 gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac
     1681 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca
     1741 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca
     1801 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca
     1861 gccagtctgc aggatatcgt cgacgaattc ggccggccgc ttcgagcaga catgataaga
     1921 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1981 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2041 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     2101 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgaccga tgcccttgag
     2161 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     2221 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     2281 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2341 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2401 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2461 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2521 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2581 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2641 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2701 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2761 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2821 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2881 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2941 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3001 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3061 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3121 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3181 aaagtatata tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg
     3241 ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat
     3301 agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc
     3361 cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa
     3421 ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca
     3481 gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag
     3541 agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt
     3601 caactctggt tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc
     3661 agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct
     3721 catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc
     3781 cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg
     3841 ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct
     3901 catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc
     3961 cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag
     4021 cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac
     4081 acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg
     4141 ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag
     4201 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     4261 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaata
//
