LOCUS       sequence-192981-        4322 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 4322)
  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 4322)
  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_OCT4 3UTR vector
FEATURES             Location/Qualifiers
     source          1..4322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          191..323
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     306..325
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        join(458..463,464..481,482..488)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             542..1201
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(608..627)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1543..1848
                     /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     1727..1746
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1926..2047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1937..1956)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1991..2010
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2292..2309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2463..3051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2543..2562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3251..4042,4043..4111))
                     /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    4216..4264
                     /note="synthetic polyadenylation signal"
     misc_feature    join(4278..4322,1..47)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     join(4318..4322,1..15)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctggc ctaactggcc
       61 ggtacctgag cattggatat tcccatcccc acctcagtaa ctgaaggaca aaccaagata
      121 agtgtgtcta tctactgtgt cccaggcttc tttatttaag aaaaaggccc ggtcgtgagg
      181 cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
      241 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
      301 ccactttgcc tttctctcca caggtgtcca ccctgcaggc ttaagcatgg ctagcaaagg
      361 agaagaactc ttcactggag ttgtcccaat tcttgttgaa ttagattgat ctagagcatg
      421 caccggtgat atcgcggccg cattaggcac cccaggcttt acactttatg cttccggctc
      481 gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa
      541 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
      601 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
      661 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
      721 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
      781 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
      841 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
      901 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
      961 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     1021 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     1081 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     1141 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     1201 aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt
     1261 ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct
     1321 atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat
     1381 atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc
     1441 gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg
     1501 aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac
     1561 acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac
     1621 acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc
     1681 tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc
     1741 gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc
     1801 gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc
     1861 cttatacaca gccagtctgc aggatatcgt cgacgaattc ggccggccgc ttcgagcaga
     1921 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1981 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2041 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2101 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgaccga
     2161 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     2221 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     2281 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2341 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2401 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2461 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2521 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2581 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2641 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2701 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2761 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2821 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2881 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2941 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3001 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3061 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gcggccgcaa atgctaaacc
     3241 actgcagtgg ttaccagtgc ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc
     3301 gttcgtccat agtggcctga ctccccgtcg tgtagatcac tacgattcgt gagggcttac
     3361 catcaggccc cagcgcagca atgatgccgc gagagccgcg ttcaccggcc cccgatttgt
     3421 cagcaatgaa ccagccagca gggagggccg agcgaagaag tggtcctgct actttgtccg
     3481 cctccatcca gtctatgagc tgctgtcgtg atgctagagt aagaagttcg ccagtgagta
     3541 gtttccgaag agttgtggcc attgctactg gcatcgtggt atcacgctcg tcgttcggta
     3601 tggcttcgtt caactctggt tcccagcggt caagccgggt cacatgatca cccatattat
     3661 gaagaaatgc agtcagctcc ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg
     3721 tgttgtcgct catggtaatg gcagcactac acaattctct taccgtcatg ccatccgtaa
     3781 gatgcttttc cgtgaccggc gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc
     3841 gaccaagctg ctcttgcccg gcgtctatac gggacaacac cgcgccacat agcagtactt
     3901 tgaaagtgct catcatcggg aatcgttctt cggggcggaa agactcaagg atcttgccgc
     3961 tattgagatc cagttcgata tagcccactc ttgcacccag ttgatcttca gcatctttta
     4021 ctttcaccag cgtttcgggg tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa
     4081 tgagtgcgac acgaaaatgt tggatgctca tactcgtcct ttttcaatat tattgaagca
     4141 tttatcaggg ttactagtac gtctctcaag gataagtaag taatattaag gtacgggagg
     4201 tattggacag gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     4261 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
     4321 gc
//
