LOCUS       sequence-192990-        4324 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 4324)
  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 4324)
  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_SULT1C vector
FEATURES             Location/Qualifiers
     source          1..4324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..108
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    288..409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(299..318)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     353..372
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(654..671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(825..1413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(905..924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1613..2404,2405..2473))
                     /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    2578..2626
                     /note="synthetic polyadenylation signal"
     misc_feature    2640..2731
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2680..2699
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     intron          2877..3009
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2992..3011
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        join(3144..3149,3150..3167,3168..3174)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3228..3887
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3294..3313)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             join(4229..4324,1..210)
                     /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"
ORIGIN
        1 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
       61 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
      121 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
      181 gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct
      241 gcaggatatc gtcgacgaat tcggccggcc gcttcgagca gacatgataa gatacattga
      301 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
      361 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
      421 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
      481 aaacctctac aaatgtggta aaatcgataa ggatccgacc gatgcccttg agagccttca
      541 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
      601 tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc
      661 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
      721 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
      781 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      841 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      901 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      961 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1021 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1081 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1141 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1201 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1261 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1321 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1381 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1441 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1501 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1561 tatgagtaaa cttggtctga cagcggccgc aaatgctaaa ccactgcagt ggttaccagt
     1621 gcttgatcag tgaggcaccg atctcagcga tctgcctatt tcgttcgtcc atagtggcct
     1681 gactccccgt cgtgtagatc actacgattc gtgagggctt accatcaggc cccagcgcag
     1741 caatgatgcc gcgagagccg cgttcaccgg cccccgattt gtcagcaatg aaccagccag
     1801 cagggagggc cgagcgaaga agtggtcctg ctactttgtc cgcctccatc cagtctatga
     1861 gctgctgtcg tgatgctaga gtaagaagtt cgccagtgag tagtttccga agagttgtgg
     1921 ccattgctac tggcatcgtg gtatcacgct cgtcgttcgg tatggcttcg ttcaactctg
     1981 gttcccagcg gtcaagccgg gtcacatgat cacccatatt atgaagaaat gcagtcagct
     2041 ccttagggcc tccgatcgtt gtcagaagta agttggccgc ggtgttgtcg ctcatggtaa
     2101 tggcagcact acacaattct cttaccgtca tgccatccgt aagatgcttt tccgtgaccg
     2161 gcgagtactc aaccaagtcg ttttgtgagt agtgtatacg gcgaccaagc tgctcttgcc
     2221 cggcgtctat acgggacaac accgcgccac atagcagtac tttgaaagtg ctcatcatcg
     2281 ggaatcgttc ttcggggcgg aaagactcaa ggatcttgcc gctattgaga tccagttcga
     2341 tatagcccac tcttgcaccc agttgatctt cagcatcttt tactttcacc agcgtttcgg
     2401 ggtgtgcaaa aacaggcaag caaaatgccg caaagaaggg aatgagtgcg acacgaaaat
     2461 gttggatgct catactcgtc ctttttcaat attattgaag catttatcag ggttactagt
     2521 acgtctctca aggataagta agtaatatta aggtacggga ggtattggac aggccgcaat
     2581 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta
     2641 acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc
     2701 agtgcaagtg caggtgccag aacatttctc tggcctaact ggccggtacc tctggtccta
     2761 gagccaacct tccatttccc cccagccccc acctgtttaa gcttcgaggc cagtgggagg
     2821 agggaggggc caggcagctg agggccagga gggcccagtc gtgaggcact gggcaggtaa
     2881 gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga
     2941 gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc
     3001 tctccacagg tgtccaccct gcaggcttaa gcatggctag caaaggagaa gaactcttca
     3061 ctggagttgt cccaattctt gttgaattag attgatctag agcatgcacc ggtgatatcg
     3121 cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
     3181 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     3241 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     3301 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     3361 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     3421 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     3481 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     3541 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     3601 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     3661 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     3721 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     3781 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     3841 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg
     3901 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     3961 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     4021 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     4081 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     4141 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     4201 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     4261 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     4321 gatg
//
