LOCUS       sequence-351765-        6278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SscINT expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6278)
  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 6278)
  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: pDY0738
            pCAG-NLS-SscINT
FEATURES             Location/Qualifiers
     source          1..6278
                     /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(3341..3358)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3347..3571
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3643..3660)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3643..3659)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3652..3674)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3800..4255)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3937..3958)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4149..4168
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4282..4386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4387..4455,4456..5247)
                     /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(4605..4624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5418..6006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5907..5926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6160..6177
                     /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 ccatgaatga gaagaacctt gagatagggg ctgcatacat tcgggtcagc
     1921 accgacgacc agactgaact gtctcccgat gctcagctgc gggtaatcct tgaggcggcc
     1981 aagaaagacg ggattataat tcctcaggag ttcgtgttca tggaggacag aggccgttcc
     2041 ggccgccggg ctgataacag acctgagttc cagaggatga tttccaccgc tcgacagaat
     2101 ccttctccat tcaggtattt atacctttgg aagttcagtc ggttcgcaag aaatcaggag
     2161 gaatcagctt tctacaaggg aattctgcgg aaaaagtgcg gcgtgacgat caaatctgtt
     2221 agtgagccca ttatggaggg catgttcggg cgcttggtag aaatgatcat cgaatggtct
     2281 gatgaattct acagcgttaa cctcagcggt gaagtcctca ggggaatgac gcaaaaggca
     2341 ttagagcatg gataccagtt aaccccctgc ctgggctacg atgctgtggg acatggaaga
     2401 ccgtacgtca tcaacgagga gcagtatcag attgttgaat ttatccaccg cagctttttc
     2461 gatggtaagg atatgacgtg gattgctagg gaagctaaca gaaggggata tcacactcgc
     2521 agggggaatc cattcgatac cagggcagtg agaatcatcc tgaccaattc tttctatgtg
     2581 ggactcgtga aatggaacga cgtaacattt caaggcacac atgagtgccg ggaaagcgtg
     2641 acttctgtat tctccgcgaa tcaggaaagg ctgaatcgta ttcaccgacc aagggggcgg
     2701 cgacaggcct cttcctgtaa acactggctg agcggcctcc tgaagtgctc aatatgcgga
     2761 gctagtctgg gctacaacca gaccaaagac ctgacaaagc gaggtcatgc tttccagtgc
     2821 tggaagtaca ccaaaggaat tcatcctggc tcttgcagcg tatcctctct caaagcagag
     2881 gcggccgttc tggagtccct gcaaatgata ttggaaactg gagaggtcga gtatacctac
     2941 gaacagcgcg agaagcacct ggatgataac aaactcaccc tcatccagaa gtccttggaa
     3001 cgacttgaca ccaaagagct gcgaattcga gaggcttacg agtctggaat agataccttg
     3061 gatgagttca agacaaataa ggcacgactg cagcgagagc gtgatcaact catggaagag
     3121 cttgaagaat tgcactctca agaggagcca gaggatgtcc ccggcaagga gatcttaatc
     3181 gaacgtattc aaaatgtata cgatttgctg caatccccag atgtcgataa tgatgataaa
     3241 ggcaacgccg tgcggtcaat tatcaagaag atagtgtata ttaaggaatc taaaactttc
     3301 tgtttttatt attatgtgtg agtcagagct cgctgatcag cctcgactgt gccttctagt
     3361 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3421 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3481 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3541 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3601 tcgagatcca ctagttctag cctcgaggct agagcggccg ccactggccg tcgttttaca
     3661 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     3721 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3781 cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3841 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3901 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3961 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4021 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4081 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4141 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4201 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4261 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4321 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4381 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4441 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4501 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4561 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4621 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4681 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4741 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4801 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4861 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4921 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4981 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5041 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5101 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5161 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5221 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5281 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5341 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5401 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5461 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5521 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5581 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5641 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5701 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5761 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5821 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5881 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5941 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6001 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6061 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6121 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6181 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6241 agctggcacg acaggtttcc cgactggaaa gcgggcag
//
