LOCUS       sequence-383161-        3131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In Situ hybridization probes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3131)
  AUTHORS   Masser EA, Noble TD, Siefert JC, Goins D, Sansam CG, Sansam CL
  TITLE     Zebrafish Rif1 impacts zygotic genome activation, replication 
            timing, and sex determination
  JOURNAL   eLife 2023 (Reviewed Preprint) 12:RP87671
REFERENCE   2  (bases 1 to 3131)
  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: "eLife 2023 
            (Reviewed Preprint) 12:RP87671"
COMMENT     Sequence Label: pUC57 beta-Globin
FEATURES             Location/Qualifiers
     source          1..3131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..22
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6..22
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        48..66
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(478..497)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(479..497)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(537..553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(537..553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(550..572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    561..577
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(585..591,592..609,610..615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    630..651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(768..785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(939..1527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1019..1038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1698..2489,2490..2558))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2321..2340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2559..2663)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2731..2749
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2787..2809)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2909..2928
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3122..3131,1..13)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgttgtaaa acgacggcca gtgaattgac gcgtattggg atggaacaat taaccctcac
       61 taaagggaac gcgcagcgat tcagaacatc tttgccaaac tcgactttga gtcggtgggc
      121 ctcgagacct tgacaagatg cttggtcgtc tatccgtgga ctcagcggta ctttggtggg
      181 tttggaaacc tatacaacac agaggcaatc atggctaacc caaaagtcaa agcgcacggt
      241 gttgtggtcc tcaagggact ggaaaaggct ctcaataaca tggacaatat caagagcacc
      301 tatgcttctc tcagcgagct tcattcagag aaactacaag tcgacccagg caatttccgg
      361 ctgttagcgg actgcctgac cgtggtgatc gccacacgca tgaggagtga gttcaccccg
      421 gacatccaag ctgcttggca gaagttcctg tctgttgttg tgtcagccct cagaaggccc
      481 tatagtgagt cgtattagtt ccatcccaat ggcgcgccga gcttggcgta atcatggtca
      541 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
      601 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
      661 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      721 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
      781 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
      841 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
      901 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      961 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1021 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1081 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1141 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1201 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1261 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1321 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1381 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1441 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1501 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1561 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1621 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1681 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1741 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1801 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1861 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1921 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1981 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2041 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2101 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2161 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2221 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2281 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2341 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2401 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2461 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2521 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2581 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2641 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     2701 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     2761 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2821 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     2881 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     2941 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     3001 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     3061 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     3121 tcccagtcac g
//
