LOCUS       sequence-192861-        6619 bp    DNA     circular SYN 14-APR-2025
DEFINITION  pUC19 with loxP-pDHFR-TS:DHFR-TS*(Pyr-R)-Stop-loxP-DHFR-TS 3' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6619)
  TITLE     Sibley lab Floxed DHFR-TS* Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6619)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pFloxed DHFR-TS*
FEATURES             Location/Qualifiers
     source          1..6619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(415..448)
                     /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(3280..3313)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4379..4395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4379..4395)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(4396..4414)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4422..4438
                     /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(4446..4452,4453..4470,4471..4476))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4491..4512
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4629..4646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4800..5388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4880..4899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5559..6350,6351..6419))
                     /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     6182..6201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6420..6524)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6592..6610
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccataact
      421 tcgtatagca tacattatac gaagttataa gcttcgccag gctgtaaatc ccgtgagtcg
      481 tcctcacaaa tcatcaagca ggtgtcctca gggagactgc ctgactgagt tatgctaatt
      541 cctttctact ttggcgtggt cacgggcgcg ccggatcctt aattaagtct agcatgtcat
      601 tcgattttca ccccccgcgt agttcctgtg tgtcattcgt tgtcgagaca actctgtccc
      661 gccccggtgc tgttccatat gcgtgacttt cccgcaattt tttcagactt tcaggaaaga
      721 caggctccgg aacgatctcg tccatgactg gtaaatccac gacaccgcaa tggcccccag
      781 cacctctatc tctcgtgcca ggggactaac gttgtatgcg tctgcgtctt gtctttttgc
      841 attcgctttc caaaaaagag agccatccgt tcccccgcac attcaacgcc gcgagtgcgg
      901 tttttgtctt ttttgagtgg taggacgctt ttcatgcgcg aactacgtgg acattaagtt
      961 ccattctctt tttcgacagc acgaaacctt gcattcaaac ccgcccgcgg aagatccgat
     1021 cttgctgctg ttcgcagtcc cagtagcgtc ctgtcggccg cgccgtctct gttggtgggc
     1081 agccgctaca cctgttatct gactgccgtg cgcgaaaatg acgccatttt tgggaaaatc
     1141 ggggaacttc attctttaaa agtatgcgga ggtttccttt ttcttctgtt cgtttctttt
     1201 tctcgggttt gataaccgtg ttcgatgtaa gcactttccg tctctcctcc gtgctttgtt
     1261 cgacatcgag accaggtgtg cagatccttc gcttgtcgat ccggagacgc gtgtctcgta
     1321 gaaccttttc attttaccac acggcagtgc ggagcactgc tctgagtgca gcagggacgg
     1381 gtgaagtttc gctttagtag tgcgtttctg ctctacgggg cgttgtcgtg tctgggaaga
     1441 tgcagaaacc ggtgtgtctg gtcgtcgcga tgacccccaa gaggggcatc ggcatcaaca
     1501 acggcctccc gtggccccac ttgaccacag atttcaaaca ctttcgtcgt gtgacaaaaa
     1561 cgacgcccga agaagccagt cgcctgaacg ggtggcttcc caggaaattt gcaaagacgg
     1621 gcgactctgg acttccctct ccatcagtcg gcaagagatt caacgccgtt gtcatgggac
     1681 ggaaaaactg ggaaagcatg cctcgaaagt ttagacccct cgtggacaga ttgaacatcg
     1741 tcgtttcctc ttccctcaaa gaagaagaca ttgcggcgga gaagcctcaa gctgaaggcc
     1801 agcagcgcgt ccgagtctgt gcttcactcc cagcagctct cagccttctg gaggaagagt
     1861 acaaggattc tgtcgaccag atttttgtcg tgggaggagc gggactgtac gaggcagcgc
     1921 tgtctctggg cgttgcctct cacctgtaca tcacgcgtgt agcccgcgag tttccgtgcg
     1981 acgttttctt ccctgcgttc cccggagatg acattctttc aaacaaatca actgctgcgc
     2041 aggctgcagc tcctgccgag tctgtgttcg ttcccttttg tccggagctc ggaagagaga
     2101 aggacaatga agcgacgtat cgacccatct tcatttccaa gaccttctca gacaacgggg
     2161 taccctacga ctttgtggtt ctcgagaaga gaaggaagac tgacgacgca gccactgcgg
     2221 aaccgagcaa cgcaatgagc tccttgacgt ccacgaggga gacaactccc gtgcacgggt
     2281 tgcaggctcc ttcttcggcc gcagccattg ccccggtgtt ggcgtggatg gacgaagaag
     2341 accggaaaaa acgcgagcaa aaggaactga ttcgggccgt tccgcatgtt cactttagag
     2401 gccatgaaga attccagtac cttgatctca ttgccgacat tattaacaat ggaaggacaa
     2461 tggatgaccg aacgggcgtt ggtgtcatct ccaaattcgg ctgcactatg cgctactcgc
     2521 tggatcaggc ctttccactt ctcaccacaa agcgtgtgtt ctggaaaggg gtcctcgaag
     2581 agttgctgtg gttcattcgc ggcgacacga acgcaaacca tctttctgag aagggcgtga
     2641 agatctggga caagaatgtg acacgcgagt tcctcgattc gcgcaatctc ccccaccgag
     2701 aggtcggaga catcggcccg ggctacggct tccagtggag acacttcggc gcggcataca
     2761 aagacatgca cacagactac acagggcagg gcgtcgacca gctgaagaat gtgatccaga
     2821 tgctgagaac gaatccaaca gatcgtcgca tgctcatgac tgcctggaat cctgcagcgc
     2881 tggacgaaat ggcgctgccg ccttgtcact tgttgtgcca gttctacgtg aacgaccaga
     2941 aggagctgtc gtgcatcatg tatcagcggt cgtgcgatgt cggcctcggc gtccccttca
     3001 acatcgcttc ctattcgctt ttgacgctca tggttgcaca cgtctgcaac ctaaaaccta
     3061 aggagttcat tcacttcatg gggaacacgc atgtctacac gaaccatgtc gaggctttaa
     3121 aagagcagct gcggagagaa ccgagaccgt tccccattgt gaacatcctc aacaaggaac
     3181 gcatcaagga aatcgacgat ttcaccgccg aggattttga ggtcgtgggc tacgtcccgc
     3241 acggacgaat ccagatggag atggctgtct aagctgagca taacttcgta tagcatacat
     3301 tatacgaagt tatcggaaat acagaagctg cccgtctctc gttttcctct cttttcggag
     3361 ggatcaggga gagtgcctcg ggtcggagag agctgacgag ggggtgccag agacccctgt
     3421 gtcctttatc gaagaaaagg gatgactctt catgtggcat ttcacacagt ctcacctcgc
     3481 cttgttttct ttttgtcaat cagaacgaaa gcgagttgcg ggtgacgcag atgtgcgtgt
     3541 atccactcgt gaatgcgtta tcgttctgta tgccgctaga gtgctggact gttgctgtct
     3601 gcccacgaca gcagacaact ttccttctat gcacttgcag gatggtgcag cgcaaacgac
     3661 ggagagaaag gagcaccctc tcagtttccc tacgatgtgc tgtcagtttc gactcttcac
     3721 cgcgaacgat tggcgatacg tctctgttga cttgttaggc tccgaccacg aagctccctt
     3781 aactaaataa gccgcgacac ctaagtgtac accatttgca gatcgataat ctgcgaccgc
     3841 tgaatccgtc cagatcagta aaaccgcacc acctaagtgt aaaccttgtt taggtcgata
     3901 aaatgctacc aacccccacc cacaatcgag ccttgagcgt ttctgcgcac gcgttggcct
     3961 acgtgacttg ctgatgcctg cctctggcca ttcatgccag tcagtgcgca taaaaatgtg
     4021 gacacagtcg gttgacaagt gttctggcag gctacagtga cacggggaaa tcgtttctga
     4081 gagagcacca aaacctttta gctgtcacca tcttcttcgc ttgctacgtt gtctgatttg
     4141 tcgtttttcc tctcggtgtg aagcgttttt cgtaacgtca atcgacccag gacactgctt
     4201 ccagctgtct gtgccgtcct gtcgtgacca gtttgtcgcc gcgactttat gagctttttc
     4261 ctgttgagaa agcggttcgg cggccgccac gtgacatgcg ggagaagaaa acgtgaccga
     4321 acaaatcaac tggtgatcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt
     4381 catagctgtt tcctgcctat agtgagtcgt attatgtgaa attgttatcc gctcacaatt
     4441 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4501 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4561 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4621 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4681 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4741 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5221 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5461 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5521 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5581 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5641 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5701 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5761 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5821 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5881 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5941 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6001 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6061 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6121 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6181 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6241 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6301 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6361 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6421 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6481 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6541 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6601 atcacgaggc cctttcgtc
//
