LOCUS       sequence-351791-        6437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BceINT expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6437)
  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 6437)
  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: pDY0750 pCAG-NLS-BceINT
FEATURES             Location/Qualifiers
     source          1..6437
                     /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"
     CDS             1876..1896
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1897..1923
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3500..3517)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3506..3730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3802..3819)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3802..3818)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3811..3833)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3959..4414)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4096..4117)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4308..4327
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4441..4545
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4546..4614,4615..5406)
                     /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(4764..4783)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5577..6165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6066..6085
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6319..6336
                     /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 sggggcgagg 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 ccatgcccaa gaagaaacgc aaggtatacc cttatgacgt acctgattac
     1921 gccggtagct acaggccaga atccctcgac gtatgcattt accttcgcaa atccaggaag
     1981 gacgttgaag aagaacgccg cgcaatcgaa gaaggcagct cctacaacgc actggaacgg
     2041 catcggaagc gattgtttgc cattgccaag gcagaaaatc acaacatcat cgatattttt
     2101 gaagaagttg ccagtggaga gagcatacag gaaagacccc aaatgcagca gctgctcagg
     2161 aagttggaag gcaatgaaat tgatggcgtg ctggtgattg atctcgatag actcgggcgg
     2221 ggcgatatgc tggatgcggg aatgatcgat cgtgcattca gatactcatc taccaaaatt
     2281 atcaccccaa cagatgtcta cgatcctgat gacgaaagtt gggagctggt gttcgggatt
     2341 aagagtttaa tcagccgaca ggagctcaag tccatcacca aacgactgca gaatggccgg
     2401 atcgattcag tgaaggaggg gaagcacatt ggcaagaagc caccttatgg ctacttgaag
     2461 gatgagaatc tgaggctgta tccagatcca gaaaaggcct ggattgtgaa gaagattttt
     2521 gaactgatgt gtgacggaaa gggacggcag atgattgcgg ctgagttgga cagactgggt
     2581 attgaccccc ctgtgacgaa aaggggagca tgggactcta gtaccatcac cagtattata
     2641 aagaacgaag tttatacagg cgtcattgtc tgggggaaat ttaagcataa aaagaggaat
     2701 ggtaagtata cgcggcataa gaacccacag gagaagtgga ttatgtacga gaacgcccat
     2761 gaacccatta tatccaaaga gcttttcgat gcggcaaacg aagcccatag ctccagacac
     2821 aagcccgctg tcataacgag taaaaagctg actaacccac tggctggcat cttgaagtgc
     2881 aagttgtgtg gctacacaat gctcatacag actcggaagg acaggcctca taactactta
     2941 cgatgtaaca atccagcctg taagggcaag caaaaacagt cagttttcaa tttagtggag
     3001 gagaagttgc tctattcact gcagcaaatc gtggacgagt accaggccca gaaagttgaa
     3061 gaggtcgaaa ttgatgattc taaactcatc tcttttaagg aaaaggcaat aatctccaaa
     3121 gagaaggagc ttaaggagtt acaagctcag aaaggcaacc tgcatgacct gctcgaacaa
     3181 ggtatttaca cggtcgaaat cttcctggaa cggcagaaga atttggtgga aagaataacc
     3241 agcatcgaga acgacatcga ggtgctgcag aaggagattg aaactgagca gatcaaagaa
     3301 cacaataaga ccgagttcat ccccgcctta aaaacggtga tcgaatcata tcacaaaaca
     3361 accaatattg aactcaaaaa ccagctgctg aagaccattc tgagcaccgt gacatactat
     3421 aggcatcccg actggaaaac caatgaattt gaaatccagg tgtacttcaa aatctcctag
     3481 gtcagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3541 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3601 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3661 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3721 ggctctatgg cttctgaggc ggaaagaacc agctggggct cgagatccac tagttctagc
     3781 ctcgaggcta gagcggccgc cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     3841 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     3901 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggaa
     3961 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4021 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4081 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4141 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4201 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4261 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4321 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4381 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     4441 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4501 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4561 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4621 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4681 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4741 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4801 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4861 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4921 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4981 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5041 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5101 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5161 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5221 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5281 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5341 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5401 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5461 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5521 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5581 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5641 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5701 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5761 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5821 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5881 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5941 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6001 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6061 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6121 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6181 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6241 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6301 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6361 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6421 gactggaaag cgggcag
//
