LOCUS       sequence-193016-        5490 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 5490)
  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 5490)
  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 1
FEATURES             Location/Qualifiers
     source          1..5490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          175..307
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     290..309
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        join(442..447,448..465,466..472)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             526..1185
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(592..611)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1527..1832
                     /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     1711..1730
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1910..2031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1921..1940)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1975..1994
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2276..2293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2447..3035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2527..2546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3235..4026,4027..4095))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     intron          4162..4734
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(5026..5043)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5032..5256
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    5372..5420
                     /note="synthetic polyadenylation signal"
     misc_feature    join(5434..5490,1..35)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     join(5474..5490,1..3)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ccccagtgca agtgcaggtg ccagaacatt tctctggcct aactggccgg tacctgagct
       61 ccctagggta cttatataag ggggtggggg cgcgttcgtc ctcagtcgcg atcgaacact
      121 cgagccgagc agacgtgcct acggaccggg gcccggtcgt gaggcactgg gcaggtaagt
      181 atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga
      241 agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc
      301 tccacaggtg tccaccctgc aggcttaagc atggctagca aaggagaaga actcttcact
      361 ggagttgtcc caattcttgt tgaattagat tgatctagag catgcaccgg tgatatcgcg
      421 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
      481 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
      541 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
      601 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
      661 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
      721 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
      781 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
      841 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
      901 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
      961 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     1021 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     1081 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     1141 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc
     1201 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     1261 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
     1321 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     1381 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     1441 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     1501 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     1561 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     1621 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     1681 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     1741 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     1801 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     1861 ctgcaggata tcgtcgacga attcggccgg ccgcttcgag cagacatgat aagatacatt
     1921 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1981 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2041 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2101 taaaacctct acaaatgtgg taaaatcgat aaggatccga ccgatgccct tgagagcctt
     2161 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     2221 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     2281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2881 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3001 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3061 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3121 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3181 tatatgagta aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca
     3241 gtgcttgatc agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc
     3301 ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc
     3361 agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc
     3421 agcagggagg gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat
     3481 gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt
     3541 ggccattgct actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc
     3601 tggttcccag cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag
     3661 ctccttaggg cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt
     3721 aatggcagca ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac
     3781 cggcgagtac tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg
     3841 cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat
     3901 cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc
     3961 gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc
     4021 ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa
     4081 atgttggatg ctcatactcg tcctttttca atattattga agcatttatc agggttacta
     4141 gtacgcgttg agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct
     4201 attgtaaaat tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga
     4261 agatgtccct tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc
     4321 tgttgacaac cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac
     4381 tttagcttgc atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag
     4441 tactttctct aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca
     4501 cagttttaga gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat
     4561 tctggctggc gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg
     4621 cctttctctt tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag
     4681 tccaaaccgg gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg
     4741 ggcaacgtgc tggttgttgt gctgtctcat cattttggca aaggattcac tcctcaggtg
     4801 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     4861 tgagatcgtt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     4921 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa tttctcgatc
     4981 gtctagcttc agcgagggat caattctcta gagctcgctg atcagcctcg actgtgcctt
     5041 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5101 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     5161 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     5221 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     5281 ggggctcgat cctctagagt cgcctcgaac tagtacgtct ctcaaggata agtaagtaat
     5341 attaaggtac gggaggtatt ggacaggccg caataaaata tctttatttt cattacatct
     5401 gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac
     5461 gaaacaaaac aaactagcaa aataggctgt
//
