LOCUS       sequence-234967-        4083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct with scrambled-1 sgRNA sequence for microinjections in 
            zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4083)
  AUTHORS   Chong-Morrison V, Mayes S, Simoes FC, Senanayake U, Carroll DS, 
            Riley PR, Wilson SW, Sauka-Spengler T
  TITLE     Ac/Ds transposition for CRISPR/dCas9-SID4x epigenome modulation in 
            zebrafish.
  JOURNAL   Biol Open. 2023 Jun 15;12(6):bio059995. doi: 10.1242/bio.059995. 
            Epub 2023 Jun 27.
  PUBMED    37367831
REFERENCE   2  (bases 1 to 4083)
  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: "Biol 
            Open."; date: "2023-06-15"; volume: "12(6):bio059995. doi"; pages: "
            10.1242/bio.059995. Epub 2023 Jun 27"
COMMENT     Sequence Label: pVC-Ds-DrU6a:scrambled1-sgRNA-Ds
FEATURES             Location/Qualifiers
     source          1..4083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..57
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     49..66
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     50..66
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     76..95
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        76..94
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1345..1365)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1345..1363)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1384..1400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1384..1400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1397..1419)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1408..1424
                     /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(1432..1438,1439..1456,1457..1462))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1477..1498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1615..1632)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1786..2374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1866..1885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2545..3336,3337..3405))
                     /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     3168..3187
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3406..3510)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3536..3991)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3673..3694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3885..3904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
       61 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacccg acgcagggat
      121 gaaagtagga tgggaaaatc ccgtaccgac cgttatcgta taaccgattt tgttagtttt
      181 atcccgatcg atttcgaacc cgaggtaaaa aacgaaaacg gaacggaaac gggatataca
      241 aaacggtaaa cggaaacgga aacggtagag ctagtttccc gaccgtttca ccgggatccc
      301 gtttttaatc gggatgatcc cgtttcgtta ccgtattttc taattcggga tgactgcaac
      361 agatccctcg actcgagttt acgtatttct ccagcctcgg tcattcagta agacgagttt
      421 cctgctctcg gcgtcttttg ttctggtctt caaggagggg ggaaatgttc tgcgcatgcc
      481 tgtgggggag ggagaaggac acgtcactga aaacgtccct gcatcacacc gagacaccca
      541 atcactcaag ccaagaccag ataattttgc atatgcttta cagtttgaaa attaccacgg
      601 taatccctca cacaaactct ggatttgaga tctttcaggt tttatcagtt tgcaggttta
      661 tgtcaccatg atatagggtc agacttgatc taagggagct gaataagtgg tttagtcact
      721 caccacctcc caaaaacata cccagaagtc cctggtatat atagctctcc ctccagctct
      781 tggttcgagc gcctacgcgc atggcctgtt taagagctat gctggaaaca gcatagcaag
      841 tttaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
      901 ctagcaagct tgcggcgaag cagatatcga attcccatgg tctagaagta gacttatatg
      961 gcttcttatg ttagccaaga gcccaagact tatcacttat gtgctacatt aaactatgtg
     1021 tgctccagat ttatatggat tttatctatg tttaattaag acttgtgttt acaatttttt
     1081 atatttgttt ttaagttttg aatatatgtt ttcatgtgtg attttaccga acaaaaatac
     1141 cggttcccgt ccgatttcga ctttaacccg accggatcgt atcggttttc gattaccgta
     1201 tttatcccgt tcgttttcgt taccggtata tcccgttttc gtttccgtcc cgcaagttaa
     1261 atatgaaaat gaaaacggta gaggtatttt accgaccgtt accgaccgtt ttcatcccta
     1321 gtcggggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     1381 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1441 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1501 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1561 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1621 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1681 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1741 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1981 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2221 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2461 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2521 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2581 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2641 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2701 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2761 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2821 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2881 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2941 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3001 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3061 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3121 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3181 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3241 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3301 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3361 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3421 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3481 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     3541 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3601 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3661 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3721 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3781 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3841 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3901 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3961 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     4021 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     4081 gct
//
