LOCUS       sequence-442662-        6316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TP901-1 Integrase expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6316)
  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 6316)
  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: pDY0310 TP901-1 Integrase
FEATURES             Location/Qualifiers
     source          1..6316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..658
                     /label=chicken beta-actin promoter
     intron          659..1675
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1599..1621
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1688..1706
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1732..1752
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3230..3247)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3236..3460
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3532..3549)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3532..3548)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3541..3563)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3689..4144)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3826..3847)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4038..4057
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4171..4275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4276..4344,4345..5136)
                     /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(4494..4513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5307..5895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5796..5815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6049..6066
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6183..6204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6219..6224,6225..6242,6243..6249)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6257..6273
                     /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     6262..6284
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6281..6297
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6281..6297
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      841 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
      901 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
     1021 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
     1081 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
     1141 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
     1201 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     1261 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
     1321 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
     1381 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     1441 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
     1561 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1621 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagatcct
     1681 taattaataa tacgactcac tatagggggt cgacccgcca ccatgacagc gccaaagaaa
     1741 aagaggaagg tcatgaccaa gaaagtggcc atctatacta gagtgagcac aacgaatcag
     1801 gccgaggagg ggttctctat tgacgagcaa atcgatcgtc tgaccaagta cgcggaagca
     1861 atgggctggc aagtcagcga cacttacacc gatgctgggt tctccggcgc caaactggaa
     1921 aggcctgcca tgcagcggct gattaacgac attgagaaca aggcctttga tacagtgctc
     1981 gtatacaagc tcgacaggct cagccgatct gtgcgggaca cgctttacct cgtaaaggat
     2041 gttttcacta agaataaaat cgacttcatt agcctgaacg aatccattga caccagctca
     2101 gctatgggct ctctgttcct gaccatcctg agcgctatca atgagtttga gagggagaat
     2161 ataaaggagc gcatgacaat gggaaagctg ggtagagcga agtccgggaa atctatgatg
     2221 tggaccaaga ccgcttttgg atactaccac aataggaaga cgggcattct ggagatcgtg
     2281 cccttgcagg caaccatcgt tgagcagatc ttcaccgact acctgagcgg aatatctctc
     2341 acgaagttgc gagataagct gaatgagagc ggacacattg gcaaggatat tccttggtca
     2401 tatagaaccc tccgccaaac tctggataat ccggtgtact gcggttacat caagttcaaa
     2461 gacagcctct tcgagggaat gcataaacct atcattccat acgagacata cctgaaagtc
     2521 caaaaggaac tcgaagagcg ccagcaacag acttacgaac ggaataataa tcccaggcct
     2581 ttccaggcca aatatatgct gtccggcatg gcaagatgcg gatactgcgg ggcaccactc
     2641 aagattgtgc ttggccataa acggaaggat ggaagcagaa ccatgaaata tcactgcgca
     2701 aaccgctttc caaggaaaac gaaggggatt accgtgtaca atgacaacaa aaaatgtgat
     2761 agcggaacct acgatctgtc caacttggaa aacaccgtca ttgacaattt aattggattt
     2821 caggaaaata atgacagcct tctgaagatt atcaacggga acaatcagcc gattctggac
     2881 acttcatctt tcaaaaaaca gatctctcag attgataaga aaattcagaa aaattccgat
     2941 ttatacctca atgatttcat aacgatggat gagctgaagg accggaccga cagtttgcag
     3001 gccgagaaga aactgctgaa agcaaagatc tccgagaaca agttcaatga cagtaccgat
     3061 gtcttcgagt tggtgaagac ccagctgggt agtatcccaa tcaacgagtt gagctatgac
     3121 aataagaaga agattgttaa taacctggtg agcaaagtgg acgtgaccgc tgataacgtg
     3181 gatattatct tcaagttcca gctggcctga gtcagagctc gctgatcagc ctcgactgtg
     3241 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3301 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3361 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3421 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3481 agctggggct cgagatccac tagttctagc ctcgaggcta gagcggccgc cactggccgt
     3541 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     3601 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3661 acagttgcgc agcctgaatg gcgaatggaa attgtaagcg ttaatatttt gttaaaattc
     3721 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3781 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3841 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3901 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3961 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4021 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4081 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4141 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4201 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4261 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4321 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4381 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4441 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4501 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4561 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4621 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4681 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4741 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4801 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4861 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4921 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4981 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5041 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5101 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5161 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5221 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5281 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5341 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5401 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5461 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5521 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5581 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5641 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5701 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5761 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5821 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5881 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5941 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6001 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6061 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6121 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6181 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     6241 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     6301 gaggcgcgcc ggattc
//
