LOCUS       sequence-183833-        4274 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 4274)
  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 4274)
  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_mCP vector
FEATURES             Location/Qualifiers
     source          1..4274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        39..69
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     intron          96..228
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     211..230
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        join(363..368,369..386,387..393)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             447..1106
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(513..532)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1448..1753
                     /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     1632..1651
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1831..1952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1842..1861)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1896..1915
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2197..2214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2368..2956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2448..2467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3156..3947,3948..4016))
                     /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    4121..4169
                     /note="synthetic polyadenylation signal"
     misc_feature    4183..4274
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4223..4242
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagag agggtatata atggaagctc
       61 gacttccagg ggcccggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg
      121 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
      181 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtccaccctg
      241 caggcttaag catggctagc aaaggagaag aactcttcac tggagttgtc ccaattcttg
      301 ttgaattaga ttgatctaga gcatgcaccg gtgatatcgc ggccgcatta ggcaccccag
      361 gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag gatccgtcga
      421 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
      481 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
      541 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
      601 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
      661 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
      721 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
      781 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
      841 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
      901 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
      961 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     1021 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     1081 gcgatgagtg gcagggcggg gcgtaaacgc gtggatccgg cttactaaaa gccagataac
     1141 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
     1201 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
     1261 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
     1321 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
     1381 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
     1441 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     1501 ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag
     1561 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     1621 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     1681 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     1741 ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggat atcgtcgacg
     1801 aattcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     1861 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1921 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1981 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2041 gtaaaatcga taaggatccg accgatgccc ttgagagcct tcaacccagt cagctccttc
     2101 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     2161 ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2221 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2281 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2341 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2401 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2461 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2521 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2581 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2641 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2701 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2761 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2821 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2881 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2941 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3001 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3061 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3121 tgacagcggc cgcaaatgct aaaccactgc agtggttacc agtgcttgat cagtgaggca
     3181 ccgatctcag cgatctgcct atttcgttcg tccatagtgg cctgactccc cgtcgtgtag
     3241 atcactacga ttcgtgaggg cttaccatca ggccccagcg cagcaatgat gccgcgagag
     3301 ccgcgttcac cggcccccga tttgtcagca atgaaccagc cagcagggag ggccgagcga
     3361 agaagtggtc ctgctacttt gtccgcctcc atccagtcta tgagctgctg tcgtgatgct
     3421 agagtaagaa gttcgccagt gagtagtttc cgaagagttg tggccattgc tactggcatc
     3481 gtggtatcac gctcgtcgtt cggtatggct tcgttcaact ctggttccca gcggtcaagc
     3541 cgggtcacat gatcacccat attatgaaga aatgcagtca gctccttagg gcctccgatc
     3601 gttgtcagaa gtaagttggc cgcggtgttg tcgctcatgg taatggcagc actacacaat
     3661 tctcttaccg tcatgccatc cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag
     3721 tcgttttgtg agtagtgtat acggcgacca agctgctctt gcccggcgtc tatacgggac
     3781 aacaccgcgc cacatagcag tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg
     3841 cggaaagact caaggatctt gccgctattg agatccagtt cgatatagcc cactcttgca
     3901 cccagttgat cttcagcatc ttttactttc accagcgttt cggggtgtgc aaaaacaggc
     3961 aagcaaaatg ccgcaaagaa gggaatgagt gcgacacgaa aatgttggat gctcatactc
     4021 gtcctttttc aatattattg aagcatttat cagggttact agtacgtctc tcaaggataa
     4081 gtaagtaata ttaaggtacg ggaggtattg gacaggccgc aataaaatat ctttattttc
     4141 attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca
     4201 aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc
     4261 cagaacattt ctct
//
