LOCUS       sequence-329835-        7315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Floxed DHFR fused with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7315)
  TITLE     Sibley lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7315)
  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            "Unpublished"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: pSL2-DHFR-mCherry-loxP.dna
FEATURES             Location/Qualifiers
     source          1..7315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(37..70)
                     /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(3025..3043)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3293..3312
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3598..3631)
                     /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(4697..4713)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4697..4713)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(4714..4732)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4740..4756
                     /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(4764..4770,4771..4788,4789..4794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4809..4830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4947..4964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5118..5706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5198..5217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5877..6668,6669..6737))
                     /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     6500..6519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6738..6842)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6910..6928
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6966..6988)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7088..7107
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7301..7315,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7315,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacccataa cttcgtatag catacattat
       61 acgaagttat aagcttcgcc aggctgtaaa tcccgtgagt cgtcctcaca aatcatcaag
      121 caggtgtcct cagggagact gcctgactga gttatgctaa ttcctttcta ctttggcgtg
      181 gtcacgggcg cgccggatcc ttaattaagt ctagcatgtc attcgatttt caccccccgc
      241 gtagttcctg tgtgtcattc gttgtcgaga caactctgtc ccgccccggt gctgttccat
      301 atgcgtgact ttcccgcaat tttttcagac tttcaggaaa gacaggctcc ggaacgatct
      361 cgtccatgac tggtaaatcc acgacaccgc aatggccccc agcacctcta tctctcgtgc
      421 caggggacta acgttgtatg cgtctgcgtc ttgtcttttt gcattcgctt tccaaaaaag
      481 agagccatcc gttcccccgc acattcaacg ccgcgagtgc ggtttttgtc ttttttgagt
      541 ggtaggacgc ttttcatgcg cgaactacgt ggacattaag ttccattctc tttttcgaca
      601 gcacgaaacc ttgcattcaa acccgcccgc ggaagatccg atcttgctgc tgttcgcagt
      661 cccagtagcg tcctgtcggc cgcgccgtct ctgttggtgg gcagccgcta cacctgttat
      721 ctgactgccg tgcgcgaaaa tgacgccatt tttgggaaaa tcggggaact tcattcttta
      781 aaagtatgcg gaggtttcct ttttcttctg ttcgtttctt tttctcgggt ttgataaccg
      841 tgttcgatgt aagcactttc cgtctctcct ccgtgctttg ttcgacatcg agaccaggtg
      901 tgcagatcct tcgcttgtcg atccggagac gcgtgtctcg tagaaccttt tcattttacc
      961 acacggcagt gcggagcact gctctgagtg cagcagggac gggtgaagtt tcgctttagt
     1021 agtgcgtttc tgctctacgg ggcgttgtcg tgtctgggaa gatgcagaaa ccggtgtgtc
     1081 tggtcgtcgc gatgaccccc aagaggggca tcggcatcaa caacggcctc ccgtggcccc
     1141 acttgaccac agatttcaaa cactttcgtc gtgtgacaaa aacgacgccc gaagaagcca
     1201 gtcgcctgaa cgggtggctt cccaggaaat ttgcaaagac gggcgactct ggacttccct
     1261 ctccatcagt cggcaagaga ttcaacgccg ttgtcatggg acggaaaaac tgggaaagca
     1321 tgcctcgaaa gtttagaccc ctcgtggaca gattgaacat cgtcgtttcc tcttccctca
     1381 aagaagaaga cattgcggcg gagaagcctc aagctgaagg ccagcagcgc gtccgagtct
     1441 gtgcttcact cccagcagct ctcagccttc tggaggaaga gtacaaggat tctgtcgacc
     1501 agatttttgt cgtgggagga gcgggactgt acgaggcagc gctgtctctg ggcgttgcct
     1561 ctcacctgta catcacgcgt gtagcccgcg agtttccgtg cgacgttttc ttccctgcgt
     1621 tccccggaga tgacattctt tcaaacaaat caactgctgc gcaggctgca gctcctgccg
     1681 agtctgtgtt cgttcccttt tgtccggagc tcggaagaga gaaggacaat gaagcgacgt
     1741 atcgacccat cttcatttcc aagaccttct cagacaacgg ggtaccctac gactttgtgg
     1801 ttctcgagaa gagaaggaag actgacgacg cagccactgc ggaaccgagc aacgcaatga
     1861 gctccttgac gtccacgagg gagacaactc ccgtgcacgg gttgcaggct ccttcttcgg
     1921 ccgcagccat tgccccggtg ttggcgtgga tggacgaaga agaccggaaa aaacgcgagc
     1981 aaaaggaact gattcgggcc gttccgcatg ttcactttag aggccatgaa gaattccagt
     2041 accttgatct cattgccgac attattaaca atggaaggac aatggatgac cgaacgggcg
     2101 ttggtgtcat ctccaaattc ggctgcacta tgcgctactc gctggatcag gcctttccac
     2161 ttctcaccac aaagcgtgtg ttctggaaag gggtcctcga agagttgctg tggttcattc
     2221 gcggcgacac gaacgcaaac catctttctg agaagggcgt gaagatctgg gacaagaatg
     2281 tgacacgcga gttcctcgat tcgcgcaatc tcccccaccg agaggtcgga gacatcggcc
     2341 cgggctacgg cttccagtgg agacacttcg gcgcggcata caaagacatg cacacagact
     2401 acacagggca gggcgtcgac cagctgaaga atgtgatcca gatgctgaga acgaatccaa
     2461 cagatcgtcg catgctcatg actgcctgga atcctgcagc gctggacgaa atggcgctgc
     2521 cgccttgtca cttgttgtgc cagttctacg tgaacgacca gaaggagctg tcgtgcatca
     2581 tgtatcagcg gtcgtgcgat gtcggcctcg gcgtcccctt caacatcgct tcctattcgc
     2641 ttttgacgct catggttgca cacgtctgca acctaaaacc taaggagttc attcacttca
     2701 tggggaacac gcatgtctac acgaaccatg tcgaggcttt aaaagagcag ctgcggagag
     2761 aaccgagacc gttccccatt gtgaacatcc tcaacaagga acgcatcaag gaaatcgacg
     2821 atttcaccgc cgaggatttt gaggtcgtgg gctacgtccc gcacggacga atccagatgg
     2881 agatggctgt cggtggaggt gctagcatgg ccatcatcaa ggagttcatg cgcttcaagg
     2941 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     3001 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     3061 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     3121 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     3181 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     3241 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     3301 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     3361 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     3421 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     3481 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     3541 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa gctgagcata
     3601 acttcgtata gcatacatta tacgaagtta tcggaaatac agaagctgcc cgtctctcgt
     3661 tttcctctct tttcggaggg atcagggaga gtgcctcggg tcggagagag ctgacgaggg
     3721 ggtgccagag acccctgtgt cctttatcga agaaaaggga tgactcttca tgtggcattt
     3781 cacacagtct cacctcgcct tgttttcttt ttgtcaatca gaacgaaagc gagttgcggg
     3841 tgacgcagat gtgcgtgtat ccactcgtga atgcgttatc gttctgtatg ccgctagagt
     3901 gctggactgt tgctgtctgc ccacgacagc agacaacttt ccttctatgc acttgcagga
     3961 tggtgcagcg caaacgacgg agagaaagga gcaccctctc agtttcccta cgatgtgctg
     4021 tcagtttcga ctcttcaccg cgaacgattg gcgatacgtc tctgttgact tgttaggctc
     4081 cgaccacgaa gctcccttaa ctaaataagc cgcgacacct aagtgtacac catttgcaga
     4141 tcgataatct gcgaccgctg aatccgtcca gatcagtaaa accgcaccac ctaagtgtaa
     4201 accttgttta ggtcgataaa atgctaccaa cccccaccca caatcgagcc ttgagcgttt
     4261 ctgcgcacgc gttggcctac gtgacttgct gatgcctgcc tctggccatt catgccagtc
     4321 agtgcgcata aaaatgtgga cacagtcggt tgacaagtgt tctggcaggc tacagtgaca
     4381 cggggaaatc gtttctgaga gagcaccaaa accttttagc tgtcaccatc ttcttcgctt
     4441 gctacgttgt ctgatttgtc gtttttcctc tcggtgtgaa gcgtttttcg taacgtcaat
     4501 cgacccagga cactgcttcc agctgtctgt gccgtcctgt cgtgaccagt ttgtcgccgc
     4561 gactttatga gctttttcct gttgagaaag cggttcggcg gccgccacgt gacatgcggg
     4621 agaagaaaac gtgaccgaac aaatcaactg gtgatctaga gtcgacctgc aggcatgcaa
     4681 gcttggcgta atcatggtca tagctgtttc ctgcctatag tgagtcgtat tatgtgaaat
     4741 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4801 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4861 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4921 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4981 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5041 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5221 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5281 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5341 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5401 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5461 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5521 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5581 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5641 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5701 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5761 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5821 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5881 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5941 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6001 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6061 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6121 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6181 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6241 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6301 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6361 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6421 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6481 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6541 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6601 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6661 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6721 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6781 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6841 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6901 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     6961 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     7021 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     7081 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     7141 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     7201 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     7261 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgtt
//
