LOCUS       sequence-192862-        8039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TgUPRT disruption by knock in with Floxed DHFR-TS*(Pyr-R) cassette 
            with UPRT homology flanks..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8039)
  TITLE     Sibley lab Floxed DHFR-TS* Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8039)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pUPRT::Floxed DHFR-TS*
FEATURES             Location/Qualifiers
     source          1..8039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..38)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(21..37)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(30..52)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     365..387
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(425..443)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        511..615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(616..684,685..1476)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(834..853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1647..2235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2136..2155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2362..2384
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2381..2397
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2381..2397
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2417..2437
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    complement(3211..3244)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(6076..6109)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(join(8032..8039,1..13))
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 ttatacatag ttgataattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
       61 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
      121 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
      181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
      241 ctcagtgcaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
      301 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
      361 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
      421 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
      481 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
      541 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
      601 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
      661 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
      721 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
      781 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
      841 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
      901 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
      961 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1021 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1081 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1141 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1201 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1261 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1321 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggt cagatggtaa gccctcccgt
     1381 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     1441 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     1501 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     1561 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     1621 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1681 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1741 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1801 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1861 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1921 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1981 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2041 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2101 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2161 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2221 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggt cttttgctgg
     2281 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2341 gcctttgagt gagctgatac cagcggataa caatttcaca caggaaacag ctatgaccat
     2401 gattacgcca agctatcaac tttgtataga aaagttgagc tactcctgtg tctgagtggg
     2461 aattaggtct cccgtatgtt ttccatctct gccccgtcta tatcccctgt ttgtctcctt
     2521 cagtcttcta cgccgaccgc ctgattcgcc tcctcatcga agaagctttg aacggtgagt
     2581 ctcaagtctc aaaagcagat ccgcttctct tgtactgcta gcatgagtct gaaaatgtag
     2641 gcaacgccta aaaagcggtg catgacccac ttcagtctac gtcaaaaaca cgccgctttt
     2701 ttggagagat ggagttaatc ggtttgctga gaaactgtac aatttattta attcgagact
     2761 tggattgttt gacatggcag gaagggtctg agcgtgcagc tgatgcaggc agactctagt
     2821 gagtgtccca acgtcgcaag taactgctgg aagcatgcag ccatatacga aactccgagg
     2881 agtaaattgt gttcttctgt cggtgtctca catgtgtttt ctgtttcgag ggtcaacatg
     2941 tttacatatg tagatatgag tttgaacgcg tgtatcccgc ttcatacgta cgtctgtctc
     3001 tgattgcctt cttcacttct tcatgtctat caacatgtta ctgcgtttgc ggatgcggag
     3061 ttctagactt tcaactgacg tcgggtgcct acgttcttct acattttact gcagaactgc
     3121 cgttcgaaaa gaaggaagtg acaacccctc tggatgtgtc ataccatgga gtttccttct
     3181 attccaagat cgaattcgag ctcggtaccc ataacttcgt atagcataca ttatacgaag
     3241 ttataagctt cgccaggctg taaatcccgt gagtcgtcct cacaaatcat caagcaggtg
     3301 tcctcaggga gactgcctga ctgagttatg ctaattcctt tctactttgg cgtggtcacg
     3361 ggcgcgccgg atccttaatt aagtctagca tgtcattcga ttttcacccc ccgcgtagtt
     3421 cctgtgtgtc attcgttgtc gagacaactc tgtcccgccc cggtgctgtt ccatatgcgt
     3481 gactttcccg caattttttc agactttcag gaaagacagg ctccggaacg atctcgtcca
     3541 tgactggtaa atccacgaca ccgcaatggc ccccagcacc tctatctctc gtgccagggg
     3601 actaacgttg tatgcgtctg cgtcttgtct ttttgcattc gctttccaaa aaagagagcc
     3661 atccgttccc ccgcacattc aacgccgcga gtgcggtttt tgtctttttt gagtggtagg
     3721 acgcttttca tgcgcgaact acgtggacat taagttccat tctctttttc gacagcacga
     3781 aaccttgcat tcaaacccgc ccgcggaaga tccgatcttg ctgctgttcg cagtcccagt
     3841 agcgtcctgt cggccgcgcc gtctctgttg gtgggcagcc gctacacctg ttatctgact
     3901 gccgtgcgcg aaaatgacgc catttttggg aaaatcgggg aacttcattc tttaaaagta
     3961 tgcggaggtt tcctttttct tctgttcgtt tctttttctc gggtttgata accgtgttcg
     4021 atgtaagcac tttccgtctc tcctccgtgc tttgttcgac atcgagacca ggtgtgcaga
     4081 tccttcgctt gtcgatccgg agacgcgtgt ctcgtagaac cttttcattt taccacacgg
     4141 cagtgcggag cactgctctg agtgcagcag ggacgggtga agtttcgctt tagtagtgcg
     4201 tttctgctct acggggcgtt gtcgtgtctg ggaagatgca gaaaccggtg tgtctggtcg
     4261 tcgcgatgac ccccaagagg ggcatcggca tcaacaacgg cctcccgtgg ccccacttga
     4321 ccacagattt caaacacttt cgtcgtgtga caaaaacgac gcccgaagaa gccagtcgcc
     4381 tgaacgggtg gcttcccagg aaatttgcaa agacgggcga ctctggactt ccctctccat
     4441 cagtcggcaa gagattcaac gccgttgtca tgggacggaa aaactgggaa agcatgcctc
     4501 gaaagtttag acccctcgtg gacagattga acatcgtcgt ttcctcttcc ctcaaagaag
     4561 aagacattgc ggcggagaag cctcaagctg aaggccagca gcgcgtccga gtctgtgctt
     4621 cactcccagc agctctcagc cttctggagg aagagtacaa ggattctgtc gaccagattt
     4681 ttgtcgtggg aggagcggga ctgtacgagg cagcgctgtc tctgggcgtt gcctctcacc
     4741 tgtacatcac gcgtgtagcc cgcgagtttc cgtgcgacgt tttcttccct gcgttccccg
     4801 gagatgacat tctttcaaac aaatcaactg ctgcgcaggc tgcagctcct gccgagtctg
     4861 tgttcgttcc cttttgtccg gagctcggaa gagagaagga caatgaagcg acgtatcgac
     4921 ccatcttcat ttccaagacc ttctcagaca acggggtacc ctacgacttt gtggttctcg
     4981 agaagagaag gaagactgac gacgcagcca ctgcggaacc gagcaacgca atgagctcct
     5041 tgacgtccac gagggagaca actcccgtgc acgggttgca ggctccttct tcggccgcag
     5101 ccattgcccc ggtgttggcg tggatggacg aagaagaccg gaaaaaacgc gagcaaaagg
     5161 aactgattcg ggccgttccg catgttcact ttagaggcca tgaagaattc cagtaccttg
     5221 atctcattgc cgacattatt aacaatggaa ggacaatgga tgaccgaacg ggcgttggtg
     5281 tcatctccaa attcggctgc actatgcgct actcgctgga tcaggccttt ccacttctca
     5341 ccacaaagcg tgtgttctgg aaaggggtcc tcgaagagtt gctgtggttc attcgcggcg
     5401 acacgaacgc aaaccatctt tctgagaagg gcgtgaagat ctgggacaag aatgtgacac
     5461 gcgagttcct cgattcgcgc aatctccccc accgagaggt cggagacatc ggcccgggct
     5521 acggcttcca gtggagacac ttcggcgcgg catacaaaga catgcacaca gactacacag
     5581 ggcagggcgt cgaccagctg aagaatgtga tccagatgct gagaacgaat ccaacagatc
     5641 gtcgcatgct catgactgcc tggaatcctg cagcgctgga cgaaatggcg ctgccgcctt
     5701 gtcacttgtt gtgccagttc tacgtgaacg accagaagga gctgtcgtgc atcatgtatc
     5761 agcggtcgtg cgatgtcggc ctcggcgtcc ccttcaacat cgcttcctat tcgcttttga
     5821 cgctcatggt tgcacacgtc tgcaacctaa aacctaagga gttcattcac ttcatgggga
     5881 acacgcatgt ctacacgaac catgtcgagg ctttaaaaga gcagctgcgg agagaaccga
     5941 gaccgttccc cattgtgaac atcctcaaca aggaacgcat caaggaaatc gacgatttca
     6001 ccgccgagga ttttgaggtc gtgggctacg tcccgcacgg acgaatccag atggagatgg
     6061 ctgtctaagc tgagcataac ttcgtatagc atacattata cgaagttatc ggaaatacag
     6121 aagctgcccg tctctcgttt tcctctcttt tcggagggat cagggagagt gcctcgggtc
     6181 ggagagagct gacgaggggg tgccagagac ccctgtgtcc tttatcgaag aaaagggatg
     6241 actcttcatg tggcatttca cacagtctca cctcgccttg ttttcttttt gtcaatcaga
     6301 acgaaagcga gttgcgggtg acgcagatgt gcgtgtatcc actcgtgaat gcgttatcgt
     6361 tctgtatgcc gctagagtgc tggactgttg ctgtctgccc acgacagcag acaactttcc
     6421 ttctatgcac ttgcaggatg gtgcagcgca aacgacggag agaaaggagc accctctcag
     6481 tttccctacg atgtgctgtc agtttcgact cttcaccgcg aacgattggc gatacgtctc
     6541 tgttgacttg ttaggctccg accacgaagc tcccttaact aaataagccg cgacacctaa
     6601 gtgtacacca tttgcagatc gataatctgc gaccgctgaa tccgtccaga tcagtaaaac
     6661 cgcaccacct aagtgtaaac cttgtttagg tcgataaaat gctaccaacc cccacccaca
     6721 atcgagcctt gagcgtttct gcgcacgcgt tggcctacgt gacttgctga tgcctgcctc
     6781 tggccattca tgccagtcag tgcgcataaa aatgtggaca cagtcggttg acaagtgttc
     6841 tggcaggcta cagtgacacg gggaaatcgt ttctgagaga gcaccaaaac cttttagctg
     6901 tcaccatctt cttcgcttgc tacgttgtct gatttgtcgt ttttcctctc ggtgtgaagc
     6961 gtttttcgta acgtcaatcg acccaggaca ctgcttccag ctgtctgtgc cgtcctgtcg
     7021 tgaccagttt gtcgccgcga ctttatgagc tttttcctgt tgagaaagcg gttcggcggc
     7081 cgccacgtga catgcgggag aagaaaacgt gaccgaacaa atcaactggt gatctagagt
     7141 cgacctgcag gcgcgagtcg atggaaagcg gcttgcgggc agtttgccgc ggctgccgca
     7201 tcgggaaaat cctcatccag agagacgaaa caactgcgga gcctaaggta cggacttgtt
     7261 tgcgttctcg ctacctgggg accacagaca tcaggtgtct tcgagtgcag ctgtcgctgg
     7321 aatttcccct cctgtcgttc accgcgagaa aaagctaagg aagagggaga aggtgaagag
     7381 cgataggcaa cccaagaggc tgctgtccac tttgttacac taaaacagat tggagtataa
     7441 gaagcacaac ttggcctgaa acacagaaca gagcggagag actacgcagt tgtgtctgtg
     7501 gaggaagagc acaattctgt tgtgacggga aggaagaaga gagagggact cgtaggtcag
     7561 ccggaaggga aggcaggtcg tggaaactat ggaggcaaga ggaaagaatt gctgcgctct
     7621 gactcactgc acgtgttttc cttacccatt gcctcccatt ttctgacacg gatgatattc
     7681 atcgctgaac ttgacagttg gttgtggctg tttgttgctg gtttgcgtgg ccaagaccgc
     7741 cgtgggcgcc cctctgctct tgtccgctgc acctgcttcc tccgcccttg atgtttgctc
     7801 tgttccagct gatctacgag aagctgcctg ccgacattag agatcgctgg gtgatgctgc
     7861 tcgatccgat gtgcgcgacg gcgggcagtg tgtgcaaagc gatcgaggtc ctcctgaggc
     7921 tcggcgtgaa ggaagagaga atcattttcg tcaacatttt ggctgctccc caaggcattg
     7981 aacgtgtttt caaggaatac ccgaaagtcc gcatggtcac tgctgcgtaa gcaacttta
//
