LOCUS       sequence-412536-        5562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ac transposase with SP6 promoter for in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5562)
  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 5562)
  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: pAC-SP6
FEATURES             Location/Qualifiers
     source          1..5562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(26..817,818..886))
                     /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     649..668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(887..991)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1059..1077
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1115..1137)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1237..1256
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1470..1489
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1779..1797
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1779..1796
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     5'UTR           1809..1851
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      1859..1868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     3'UTR           4201..4326
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(4427..4443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4427..4443)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4440..4462)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4451..4467
                     /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(4475..4481,4482..4499,4500..4505))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4520..4541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4658..4675)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4829..5417)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4909..4928)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
       61 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
      121 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
      181 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
      241 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
      301 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
      361 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
      421 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
      481 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
      541 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
      601 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
      661 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
      721 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
      781 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
      841 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
      901 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
      961 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     1021 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     1081 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     1141 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     1201 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     1261 gagtgcacca ttcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     1321 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     1381 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     1441 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     1501 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctggctagcg
     1561 atgaccctgc tgattggttc gctgaccatt tccgggtgcg ggacggcgtt accagaaact
     1621 cagaaggttc gtccaaccaa accgactctg acggcagttt acgagagaga tgatagggtc
     1681 tgcttcagta agccagatgc tacacaatta ggcttgtaca tattgtcgtt agaacgcggc
     1741 tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaata
     1801 caagcttgct tgttcttttt gcagaagctc agaataaacg ctcaactttg gcagatccgc
     1861 caccatgggt cctccagaga agaagcgtaa ggtagaaatg gctattgttc atgaaccaca
     1921 accacaacca caaccacaac cagaaccaca accacagcca caacctgaac ccgaagaaga
     1981 agcaccacag aagagggcaa agaagtgcac atcggatgta tggcagcatt tcaccaagaa
     2041 ggaaattgaa gtggaggtcg atggaaagaa atacgttcag gtatgggggc attgcaactt
     2101 tcctaattgc aaggctaagt atagggctga gggtcatcat ggaacaagcg gatttcgaaa
     2161 tcacttgaga acatcacata gtttagttaa aggtcagttg tgtctaaaaa gtgaaaagga
     2221 tcatggcaaa gacataaatc tcattgagcc ttataagtac gatgaagtgg ttagcctaaa
     2281 gaagcttcat ttggcaataa tcatgcatga ataccctttc aatattgtag aacatgagta
     2341 ctttgttgag tttgttaagt ctctgcgccc tcactttcca ataaagtccc gtgtcactgc
     2401 tagaaaatat atcatggatt tgtatttgga agaaaaagaa aagttgtatg gaaaactaaa
     2461 agatgttcag tctcgcttca gtacaactat ggatatgtgg acatcttgtc aaaataagtc
     2521 atacatgtgt gtcaccatcc attggattga tgatgattgg tgtctccaaa aaagaattgt
     2581 tggctttttt catgttgaag ggcgccacac tggccaaagg ttatcacaaa ccttcactgc
     2641 aatcatggtt aagtggaaca ttgagaaaaa attgtttgcc ttgtctttgg ataatgctag
     2701 tgcaaatgaa gtagctgtgc acgatataat tgaggatttg caggacactg attcaaatct
     2761 agtttgtgat ggtgctttct ttcatgtgag gtgtgcttgt cacatactga acttggttgc
     2821 aaaggatggc ttggctgtaa ttgcaggaac aattgagaaa atcaaagcga ttgttcttgc
     2881 tgtaaaatct tctcctttgc agtgggaaga actaatgaag tgtgctagtg aatgtgactt
     2941 ggataaatct aaagggatct catatgatgt ctcaactaga tggaattcaa cctatttgat
     3001 gttgagggat gccttatatt ataagcctgc actaataagg cttaaaacaa gtgatcctcg
     3061 caggtacgat gcaatttgtc ctaaagccga ggagtggaag atggcattaa ctctttttaa
     3121 gtgtttgaag aagttttttg atctcactga actcctatct ggtactcaat attccactgc
     3181 aaatttattt tacaaaggtt tctgtgagat aaaggatttg attgaccaat ggtgtgttca
     3241 tgaaaaattt gtcattagga gaatggccgt tgcaatgagt gaaaagtttg agaaatattg
     3301 gaaagtgtct aatattgcac tagctgtagc atgcttcctt gaccctaggt acaagaaaat
     3361 attgattgag ttctatatga aaaaatttca tggtgattca tacaaagttc atgtagatga
     3421 ctttgttagg gtcattagaa aattgtatca attctattct agttgtagtc cttcagctcc
     3481 aaagacaaag acaactacta atgatagtat ggatgatacc ttgatggaaa atgaagatga
     3541 tgaatttcaa aactatttgc atgagttgaa ggattatgat caagtagagt caaatgaatt
     3601 ggataaatat atgtctgaac cccttttgaa gcatagtggt cagtttgata ttttatcatg
     3661 gtggagggga agggttgcag aatatcctat tctcacccaa attgcaaggg atgtgctagc
     3721 aatacaagtg tcaactgttg cttctgagtc tgcgttcagt gctggtggtc gtgttgttga
     3781 tccttaccgc aatcgtcttg gttcggagat tgttgaagct ttgatatgca caaaagattg
     3841 ggtagcagca tctagaaaag gtgctacata ttttccaaca atgattggtg atctcgaggt
     3901 gctagactct gttattgctg ctgcaacaaa tcatgagaat catatggatg aggatgaaga
     3961 cgcaatagaa ttttctaaga ataatgaaga tgtagcaagt ggctcctctc catgagcaat
     4021 gtgtcttatg tttgttgaca gatgagcctt ggttgtaata gtttatgcat gctaagtgct
     4081 ccagatgtga gcaagtgatt atgaatatgt gttttaaact ttatattgtg tcatgtgtgc
     4141 tagtagactt atatggcttc ttatgttagc caagggggcc ccgggatttg gttaccacta
     4201 aaccagcctc aagaacaccc gaatggagtc tctaagctac ataataccaa cttacacttt
     4261 acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc tcctaataaa aagaaagttt
     4321 cttcacattc taaaaaaaaa aaaaaaaaaa aaaaaaaaaa cccccccccc ccccccctgc
     4381 aggtcgactc tagaggatcc ccgggcgagc tcgaattcgt aatcatgtca tagctgtttc
     4441 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4501 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4561 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4621 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4681 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4741 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4801 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4861 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4921 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4981 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5041 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5101 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5161 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5221 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5281 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5341 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5401 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5461 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5521 tccttttaaa ttaaaaatga agttttaaat caatctaaag ta
//
