LOCUS       sequence-193171-        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_TTF2 vector
FEATURES             Location/Qualifiers
     source          1..4324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(266..1057,1058..1126))
                     /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    1231..1279
                     /note="synthetic polyadenylation signal"
     misc_feature    1293..1384
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1333..1352
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     intron          1530..1662
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1645..1664
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        join(1797..1802,1803..1820,1821..1827)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1881..2540
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1947..1966)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2882..3187
                     /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     3066..3085
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    3265..3386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3276..3295)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3330..3349
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3631..3648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3802..4324,1..66))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3882..3901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
       61 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      121 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      181 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagcggc
      241 cgcaaatgct aaaccactgc agtggttacc agtgcttgat cagtgaggca ccgatctcag
      301 cgatctgcct atttcgttcg tccatagtgg cctgactccc cgtcgtgtag atcactacga
      361 ttcgtgaggg cttaccatca ggccccagcg cagcaatgat gccgcgagag ccgcgttcac
      421 cggcccccga tttgtcagca atgaaccagc cagcagggag ggccgagcga agaagtggtc
      481 ctgctacttt gtccgcctcc atccagtcta tgagctgctg tcgtgatgct agagtaagaa
      541 gttcgccagt gagtagtttc cgaagagttg tggccattgc tactggcatc gtggtatcac
      601 gctcgtcgtt cggtatggct tcgttcaact ctggttccca gcggtcaagc cgggtcacat
      661 gatcacccat attatgaaga aatgcagtca gctccttagg gcctccgatc gttgtcagaa
      721 gtaagttggc cgcggtgttg tcgctcatgg taatggcagc actacacaat tctcttaccg
      781 tcatgccatc cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg
      841 agtagtgtat acggcgacca agctgctctt gcccggcgtc tatacgggac aacaccgcgc
      901 cacatagcag tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg cggaaagact
      961 caaggatctt gccgctattg agatccagtt cgatatagcc cactcttgca cccagttgat
     1021 cttcagcatc ttttactttc accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg
     1081 ccgcaaagaa gggaatgagt gcgacacgaa aatgttggat gctcatactc gtcctttttc
     1141 aatattattg aagcatttat cagggttact agtacgtctc tcaaggataa gtaagtaata
     1201 ttaaggtacg ggaggtattg gacaggccgc aataaaatat ctttattttc attacatctg
     1261 tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg
     1321 aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt
     1381 ctctggccta actggccggt accgaagcct cgcctgcaca gggcacagct gcctcttgcc
     1441 tcctcttcgc ctccacggtg gaagggctgg ggccacgggg cagagaagaa aggttatctc
     1501 tgcgggcccg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
     1561 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     1621 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac cctgcaggct
     1681 taagcatggc tagcaaagga gaagaactct tcactggagt tgtcccaatt cttgttgaat
     1741 tagattgatc tagagcatgc accggtgata tcgcggccgc attaggcacc ccaggcttta
     1801 cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt
     1861 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     1921 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     1981 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     2041 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
     2101 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     2161 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     2221 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     2281 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     2341 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     2401 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     2461 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     2521 agtggcaggg cggggcgtaa acgcgtggat ccggcttact aaaagccaga taacagtatg
     2581 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta
     2641 tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta
     2701 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag
     2761 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct
     2821 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga
     2881 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     2941 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg
     3001 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     3061 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     3121 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     3181 aatataaatg tcaggctccc ttatacacag ccagtctgca ggatatcgtc gacgaattcg
     3241 gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     3301 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3361 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3421 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     3481 tcgataagga tccgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     3541 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     3601 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4201 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4321 aacc
//
