LOCUS       sequence-440874-        3206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector from which reporter gene 3'UTRs, an enhancer, and a neutral 
            DNA spacer are cloned for assembling an insulator-seq reporter 
            library..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3206)
  AUTHORS   Tonelli A, Cousin P, Gambetta MC
  TITLE     Protocol for detecting genomic insulators in Drosophila using 
            insulator-seq, a massively parallel reporter assay.
  JOURNAL   STAR Protoc. 2024 Oct 24;5(4):103391. doi: 
            10.1016/j.xpro.2024.103391.
  PUBMED    39453817
REFERENCE   2  (bases 1 to 3206)
  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: "STAR 
            Protoc."; date: "2024-10-24"; volume: "5(4):103391. doi"; pages: " 
            10.1016/j.xpro.2024.103391"
COMMENT     Sequence Label: insulator_seq_UENU_donor
FEATURES             Location/Qualifiers
     source          1..3206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..34)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1353..1457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1458..1526,1527..2318)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1676..1695)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2798..3154
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ataacttcgt atagcataca ttatacgaag ttatctgcca ggcacatggg ttttactagt
       61 atcgattcgc gacctactcc ggaatattag gtctcagaag accctagggc tttatttgta
      121 accattataa gctgcaataa acaagttgga tcccatgatg ataaacaatg tatggtgcta
      181 atgttgcttc aacaacaatt ctgttgaact gtgttttcat gtttgccaac aagcaccttt
      241 atactcggtg gcctccccac caccaacttt tttgcactgc aaaaaaacac gcttttgcac
      301 gcgggcccat acatagtaca aactctacgt ttcgtagact attttacata aatagtctac
      361 accgttgtat acgctccaaa tacactacca cacattgaac ctttttgcag tgcaaaaaag
      421 tacgtgtcgg cagtcacgta ggccggcctt atcgggtcgc gtcctgtcac gtacgaatca
      481 cattatcgga ccggacgagt gttgtcttat cgtgacagga cgccagcttc ctgtgttgct
      541 aaccgcagcc ggacgcaact ccttatcgga acaggacgcg cctccatatc agccgcgcgt
      601 tatctcatgc gcgtgaccgg acacgaggcg cccgtcccgc ttataggtac tgaatccggt
      661 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
      721 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
      781 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
      841 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
      901 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
      961 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     1021 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     1081 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcaact
     1141 tgtttattgc agcttataat ggttacaaat aaagcttcga ggtcttcgga gacccactgc
     1201 ttgagcctag aagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgtc
     1261 accgctgagc aataactatc ataaccccta ggtgccattt cattacctct ttctccgcac
     1321 ccgacataga tctgggccaa cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1381 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1441 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1501 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1561 aagatcagtt gggtgcacgg gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1621 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1681 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1741 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1801 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1861 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1921 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1981 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2041 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2101 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2161 ccggtgagcg tgggtcccgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2221 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2281 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2341 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2401 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2461 accccgtaga ggatgaatgg cagaaattcg aaagcaaatt cgacccggtc gtcggttcag
     2521 ggcagggtcg ttaaatagcc gcttatgtct attgctggtt taccggttta ttgactaccg
     2581 gaagcagtgt gaccgtgtgc ttctcaaatg cctgaggcca gtttgctcag gctctccccg
     2641 tggaggtaat aattgacgat atgatcattt attctgcctc ccagctgaca ttcatccggg
     2701 gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc
     2761 gccctgagtg cttgcggcag cgtgaagcta attcccatgt cagccgttaa gtgttcctgt
     2821 gtcactcaaa attgctttga gaggctctaa gggcttctca gtgcgttaca tccctggctt
     2881 gttgtccaca accgttaaac cttaaaagct ttaaaagcct tatatattct tttttttctt
     2941 ataaaactta aaaccttaga ggctatttaa gttgctgatt tatattaatt ttattgttca
     3001 aacatgagag cttagtacgt gaaacatgag agcttagtac gttagccatg agagcttagt
     3061 acgttagcca tgagggttta gttcgttaaa catgagagct tagtacgtta aacatgagag
     3121 cttagtacgt gaaacatgag agcttagtac gtactatcaa caggttgaac tgctgatcaa
     3181 cagatcctct acgcggccgc ggtacc
//
