LOCUS       sequence-491733-        5997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for Crispr/Cas9 gene editing of the human NTRK1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5997)
  TITLE     deNooij lab plasmids 2025
REFERENCE   2  (bases 1 to 5997)
  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"
COMMENT     Sequence Label: pDonor NTRK1-P2A-tdTomato
FEATURES             Location/Qualifiers
     source          1..5997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1162..1617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1511..1530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1744..1766
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1758..1775
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1759..1775
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1782..1801
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1782..1800
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2700..2756
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2757..4187
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYHVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     promoter        complement(4968..4986)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5007..5023)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5007..5023)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5020..5042)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5031..5047
                     /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(5055..5061,5062..5079,5080..5085))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5100..5121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5238..5255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5409..5997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5489..5508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     1621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1681 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     1741 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg
     1801 gcgaattggg tacttttttg gcgcgccgag aagaaatgga attagtgccg cccataaccc
     1861 aagtcctcta atagggaggg gggaagaagg ggaaaagagg ctccaggccc cttctccaat
     1921 cactccctgc caccctttct cctttggatt ccttggctgc tttagcagtt cttcctagag
     1981 tctaactttg atctttcttg ctgcagtttc tttttgggag agctagtcag tcccacagag
     2041 tggtatccct agaagggaga agtaaggatt gccctcttct ttaaaatgaa agccagctat
     2101 ttttcacgcc ctttaactgc agttctgctc tattttcttt tctctctctg gagctgagag
     2161 tcagagggcc cttctcctcc tcctttcagc ccccaacact aagctgatgg attgataaat
     2221 acctcagccc ctcgccttcc tcaacccacc tggcaagtct tcttaggatc tgatcccagt
     2281 tttctggaag caatcctacc ccagcccaag cttcccagag tcgagcctta atccttctca
     2341 cttctcagtg tcagagcaga aatgaatcct ggggttgact gtgtccattc gggttattag
     2401 cagctaagaa gcccagacga gtagtgtgag ctgccttggg agcctcagtg agggcactgg
     2461 gactggcctc actctcttgc ccccagccta gtgggctttc tcctctgtct ctccggtggc
     2521 cccaggcaat cgactgcatc acgcagggac gtgagttgga gcggccacgt gcctgcccac
     2581 cagaggtcta cgccatcatg cggggctgct ggcagcggga gccccagcaa cgccacagca
     2641 tcaaggatgt gcacgcccgg ctgcaagccc tggcccaggc acctcctgtc ggatccggag
     2701 ccacgaactt cagcctgctg aagcaggctg gcgacgtgga ggagaaccct ggacctatgg
     2761 tgagcaaggg cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct
     2821 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     2881 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     2941 tgtcccccca gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg
     3001 attacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     3061 acggcggtct ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca
     3121 aggtgaagat gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca
     3181 tgggctggga ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga
     3241 tccaccaggc cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct
     3301 acatggccaa gaagcccgtg caactgcccg gctactacca cgtcgacacc aagctggaca
     3361 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc
     3421 accacctgtt cctggggcat ggcaccggca gcaccggcag cggcagctcc ggcaccgcct
     3481 cctccgagga caacaacatg gccgtcatca aagagttcat gcgcttcaag gtgcgcatgg
     3541 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     3601 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
     3661 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca
     3721 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     3781 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga
     3841 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga
     3901 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
     3961 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga
     4021 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc
     4081 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg
     4141 gccgccacca cctgttcctg tacggcatgg acgagctgta caagtaatac ctggatgtcc
     4201 tgggctaggg ggccggccca ggggctggga gtggttagcc ggaatactgg ggcctgccct
     4261 cagcatcccc catagctccc agcagcccca gggtgatctc aaagtatcta attcaccctc
     4321 agcatgtggg aagggacagg tgggggctgg gagtagagga tgttcctgct tctctaggca
     4381 aggtcccgtc atagcaatta tatttattat cccttggctg tgtctcttgc cagttattgg
     4441 gatgacgtcg ttccaggagg gaggcattgg gattcaggta gggggacagc tctctgggat
     4501 cttcccccac tcaggttccc ctcagctgcc tacccctact ccatggacct gtcactctca
     4561 cctttgacta tagtttggat cccattccca tggtcacctg gctcacctgc tggtaaggca
     4621 gcctctggtc aaacttccta gaccttgaga gcctaactgt caatctttgt agttcatgtt
     4681 caggaaggct gggctatagt ggaaggggtg gaagttagat gtactggtag gatgggagcg
     4741 ttgatggggt gtcttccttg tattggtgtg tccgggacag agcaagcacg cagcagcaga
     4801 tgagagagaa acgtgtgtgt gtgtgtgggg tgatgggggg caggaaaggg acggagctgg
     4861 actaaggatt gatggaggca ggaccccttg ttcctgcccc tctgttactc ttcttagctg
     4921 gcgcgccaaa aaaggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg
     4981 ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5041 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5101 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5161 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5221 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5281 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5341 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5401 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5461 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5521 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5581 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5641 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5701 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5761 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5821 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5881 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5941 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaa
//
