LOCUS       sequence-351788-        6311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SacINT expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6311)
  AUTHORS   Yarnall MTN, Ioannidi EI, Schmitt-Ulms C, Krajeski RN, Lim J, 
            Villiger L, Zhou W, Jiang K, Garushyants SK, Roberts N, Zhang L, 
            Vakulskas CA, Walker JA 2nd, Kadina AP, Zepeda AE, Holden K, Ma H, 
            Xie J, Gao G, Foquet L, Bial G, Donnelly SK, Miyata Y, Radiloff DR, 
            Henderson JM, Ujita A, Abudayyeh OO, Gootenberg JS
  TITLE     Drag-and-drop genome insertion of large sequences without 
            double-strand DNA cleavage using CRISPR-directed integrases.
  JOURNAL   Nat Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4.
  PUBMED    36424489
REFERENCE   2  (bases 1 to 6311)
  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 
            Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4."
COMMENT     Sequence Label: pDY0739 pCAG-NLS-SacINT
FEATURES             Location/Qualifiers
     source          1..6311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    16..37
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(52..57,58..75,76..82)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    90..106
                     /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)."
     primer_bind     95..117
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     114..130
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     114..130
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        150..529
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        531..808
                     /label=chicken beta-actin promoter
     intron          809..1825
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1749..1771
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1838..1857
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1838..1856
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3374..3391)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3380..3604
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3676..3693)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3676..3692)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3685..3707)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3833..4288)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3970..3991)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4182..4201
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4315..4419
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4420..4488,4489..5280)
                     /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     complement(4638..4657)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5451..6039
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5940..5959
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6193..6210
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
       61 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      121 cagctatgac catgaggcgc gccggattcg acattgatta ttgactagtt attaatagta
      181 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      241 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      301 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      361 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      421 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      481 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
      541 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
      601 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc
      661 rggsggggsg gggsggggsg rggggsgggg sggggsgagg cggagaggtg cggcggcagc
      721 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
      781 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
      841 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
      901 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
      961 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
     1021 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     1081 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     1141 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
     1201 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1261 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
     1321 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
     1381 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga
     1441 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1501 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
     1561 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
     1621 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     1681 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     1741 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     1801 tcatgccttc ttctttttcc tacagatcct taattaataa tacgactcac tatagggggt
     1861 cgacccgcca ccatgaagga gaaggtgagt gagagaaaaa caggcgccat ttacataaga
     1921 gtttctacgg acaaacagga agagctttca ccagacgcac agctgaggct cctcctggac
     1981 tacgctaaga aagattctat cgatgttcct aaggagtaca tcttccaaga taacggcatt
     2041 agtgggcgaa aagcgaacaa gcgccccgcg ttccagaata tgatcgcact cgcgaagtcc
     2101 aaagagcacc caatcgacac aatcattgtg tggaagttct ctcgctttgc ccggaatcag
     2161 gaggaatcaa ttgtgtacaa gagtttactc aaaaaaaaca acgtcgatgt ggtgagtgtg
     2221 tccgagcctc tgattgatgg gccatttgga agcctgattg agagaattat tgagtggatg
     2281 gacgagtatt attccattcg attgtctggc gaggtgatgc gtggtatgac tcaaaatgcc
     2341 atgcgggggc attaccagag cgatgcaccg attgggtaca catccccagg ggacaaaaag
     2401 cccccggtta taaacccgga taccgttcag attcctctga tgatcaaaga tatgttctta
     2461 agcggctcaa cccagctgca aattgccaga aagctcaacg acagtggcta taggacaaag
     2521 cgcggtaacc tgtgggacgc gagaggcgtc cggtacgtcc tggagaaccc gttttacatc
     2581 gggaaaagcc gctggaatta cacggagaga gggcgacggc tgaagccggc agatgaggtg
     2641 atatacgctg acgggaactg ggaggcactg tgggatgagg acaccttcaa ggagatccaa
     2701 aaaagattgg cactgaatat gcgcaagtcc aagtctaggg acatctcagc tgcaaaacac
     2761 tggctgagcg gtctcttaat ctgttcttcc tgcggcggaa ccctggcctt cgggggagca
     2821 cacaatatga gggggtttca atgctggaaa tactcaaagg ggttctgcag cgaatcccat
     2881 tatatcagca ccggtccaat tgagaaaatg gttctggagt acttagaggc cgtcatgcac
     2941 tcccctgcgc tgagttacac ggttatcagt agttcatccg tcgatgccag ctccaaactg
     3001 tcagacctgg agcgccaatt gcagaaaata gacgccaagg agaaacgcat caaggcagca
     3061 tacctcaacg aaatagatac actggaggag tacaaagcta ataaaacagc cttggaggaa
     3121 gaacgccgta ccgtcgagaa ggaaatcgag gagctcaccc tcagcgacgt gaaatattct
     3181 aaggaggacc ttgacaagaa aatgaagcag aatatatcag acctgctgcg ggtgctgaga
     3241 gacgaatctg ccgattacat ccagaaaggt aacatgatga gaaacgtggt cgatcatatc
     3301 gtctttaaca ggaagaatac tagcctggac gtttttctga aattagtagt gtaggtcaga
     3361 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3421 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3481 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3541 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3601 atggcttctg aggcggaaag aaccagctgg ggctcgagat ccactagttc tagcctcgag
     3661 gctagagcgg ccgccactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     3721 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3781 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta
     3841 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3901 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3961 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4021 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4081 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4141 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4201 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4261 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4321 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4381 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4441 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4501 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4561 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4621 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4681 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4741 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4801 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4861 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4921 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4981 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5041 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5101 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5161 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5221 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5281 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5341 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5401 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5461 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5521 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5581 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5641 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5701 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5761 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5821 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5881 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5941 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6001 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6061 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6121 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6181 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6241 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6301 aaagcgggca g
//
