LOCUS       sequence-193173-        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"
     CDS             complement(join(610..1401,1402..1470))
                     /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    1575..1623
                     /note="synthetic polyadenylation signal"
     misc_feature    1637..1728
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1677..1696
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1767..1797
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     intron          1824..1956
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1939..1958
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        join(2091..2096,2097..2114,2115..2121)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2175..2834
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2241..2260)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3176..3481
                     /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     3360..3379
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    3559..3680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3570..3589)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3624..3643
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3925..3942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4096..4274,1..410))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4176..4195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
       61 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      241 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      541 aatcaatcta aagtatatat gagtaaactt ggtctgacag cggccgcaaa tgctaaacca
      601 ctgcagtggt taccagtgct tgatcagtga ggcaccgatc tcagcgatct gcctatttcg
      661 ttcgtccata gtggcctgac tccccgtcgt gtagatcact acgattcgtg agggcttacc
      721 atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt tcaccggccc ccgatttgtc
      781 agcaatgaac cagccagcag ggagggccga gcgaagaagt ggtcctgcta ctttgtccgc
      841 ctccatccag tctatgagct gctgtcgtga tgctagagta agaagttcgc cagtgagtag
      901 tttccgaaga gttgtggcca ttgctactgg catcgtggta tcacgctcgt cgttcggtat
      961 ggcttcgttc aactctggtt cccagcggtc aagccgggtc acatgatcac ccatattatg
     1021 aagaaatgca gtcagctcct tagggcctcc gatcgttgtc agaagtaagt tggccgcggt
     1081 gttgtcgctc atggtaatgg cagcactaca caattctctt accgtcatgc catccgtaag
     1141 atgcttttcc gtgaccggcg agtactcaac caagtcgttt tgtgagtagt gtatacggcg
     1201 accaagctgc tcttgcccgg cgtctatacg ggacaacacc gcgccacata gcagtacttt
     1261 gaaagtgctc atcatcggga atcgttcttc ggggcggaaa gactcaagga tcttgccgct
     1321 attgagatcc agttcgatat agcccactct tgcacccagt tgatcttcag catcttttac
     1381 tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa aatgccgcaa agaagggaat
     1441 gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt tttcaatatt attgaagcat
     1501 ttatcagggt tactagtacg tctctcaagg ataagtaagt aatattaagg tacgggaggt
     1561 attggacagg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     1621 gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
     1681 caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctgg cctaactggc
     1741 cggtacctga gctcgctagc ctcgagagag ggtatataat ggaagctcga cttccagggg
     1801 cccggtcgtg aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc
     1861 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     1921 gtcttactga catccacttt gcctttctct ccacaggtgt ccaccctgca ggcttaagca
     1981 tggctagcaa aggagaagaa ctcttcactg gagttgtccc aattcttgtt gaattagatt
     2041 gatctagagc atgcaccggt gatatcgcgg ccgcattagg caccccaggc tttacacttt
     2101 atgcttccgg ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag
     2161 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     2221 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     2281 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     2341 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg
     2401 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     2461 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     2521 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     2581 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
     2641 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     2701 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     2761 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     2821 agggcggggc gtaaacgcgt ggatccggct tactaaaagc cagataacag tatgcgtatt
     2881 tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa
     2941 aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt
     3001 gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa
     3061 gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc
     3121 gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca
     3181 gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag
     3241 tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct
     3301 gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg
     3361 catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga
     3421 tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata
     3481 aatgtcaggc tcccttatac acagccagtc tgcaggatat cgtcgacgaa ttcggccggc
     3541 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3601 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3661 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3721 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     3781 aggatccgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3841 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3901 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3961 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4021 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4261 tctcccttcg ggaa
//
