LOCUS       sequence-409779-        4010 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 4010)
  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 4010)
  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_ORI vector
FEATURES             Location/Qualifiers
     source          1..4010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..69,70..861)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(1051..1639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1131..1150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     intron          1735..1867
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1850..1869
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        join(2002..2007,2008..2025,2026..2032)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2086..2745
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2152..2171)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3087..3392
                     /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     3271..3290
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    3470..3591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3481..3500)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3535..3554
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3836..3853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atgagcatcc aacattttcg tgtcgcactc attcccttct ttgcggcatt ttgcttgcct
       61 gtttttgcac accccgaaac gctggtgaaa gtaaaagatg ctgaagatca actgggtgca
      121 agagtgggct atatcgaact ggatctcaat agcggcaaga tccttgagtc tttccgcccc
      181 gaagaacgat tcccgatgat gagcactttc aaagtactgc tatgtggcgc ggtgttgtcc
      241 cgtatagacg ccgggcaaga gcagcttggt cgccgtatac actactcaca aaacgacttg
      301 gttgagtact cgccggtcac ggaaaagcat cttacggatg gcatgacggt aagagaattg
      361 tgtagtgctg ccattaccat gagcgacaac accgcggcca acttacttct gacaacgatc
      421 ggaggcccta aggagctgac tgcatttctt cataatatgg gtgatcatgt gacccggctt
      481 gaccgctggg aaccagagtt gaacgaagcc ataccgaacg acgagcgtga taccacgatg
      541 ccagtagcaa tggccacaac tcttcggaaa ctactcactg gcgaacttct tactctagca
      601 tcacgacagc agctcataga ctggatggag gcggacaaag tagcaggacc acttcttcgc
      661 tcggccctcc ctgctggctg gttcattgct gacaaatcgg gggccggtga acgcggctct
      721 cgcggcatca ttgctgcgct ggggcctgat ggtaagccct cacgaatcgt agtgatctac
      781 acgacgggga gtcaggccac tatggacgaa cgaaataggc agatcgctga gatcggtgcc
      841 tcactgatca agcactggta accactgcag tggtttagca tttgcggccg ctgtcagacc
      901 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
      961 aggtgaagat cctttttgat aatctcatga catgtgagca aaaggccagc aaaaggccag
     1021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1681 gaacgaaaac tcacgttaag ggattttggg gcccggtcgt gaggcactgg gcaggtaagt
     1741 atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga
     1801 agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc
     1861 tccacaggtg tccaccctgc aggcttaagc atggctagca aaggagaaga actcttcact
     1921 ggagttgtcc caattcttgt tgaattagat tgatctagag catgcaccgg tgatatcgcg
     1981 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
     2041 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     2101 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     2161 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     2221 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     2281 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     2341 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     2401 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     2461 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     2521 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     2581 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     2641 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     2701 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc
     2761 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     2821 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
     2881 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     2941 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     3001 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     3061 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     3121 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     3181 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     3241 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     3301 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     3361 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     3421 ctgcaggata tcgtcgacga attcggccgg ccgcttcgag cagacatgat aagatacatt
     3481 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3541 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3601 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     3661 taaaacctct acaaatgtgg taaaatcgat aaggatccga ccgatgccct tgagagcctt
     3721 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     3781 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     3841 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3901 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tcttgagaga
     3961 cgtactagta accctgataa atgcttcaat aatattgaaa aaggacgagt
//
