LOCUS       sequence-10394-e        5134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5134)
  AUTHORS   Chen SX, Osipovich AB, Ustione A, Potter LA, Hipkens S, Gangula R, 
            Yuan W, Piston DW, Magnuson MA
  TITLE     Quantification of factors influencing fluorescent protein expression
            using RMCE to generate an allelic series in the ROSA26 locus in 
            mice.
  JOURNAL   Dis Model Mech. 2011 Jul;4(4):537-47. doi: 10.1242/dmm.006569. Epub 
            2011 Feb 14.
  PUBMED    21324933
REFERENCE   2  (bases 1 to 5134)
  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"; doi: 
            "10.1242/dmm.006569"; journalName: "Dis Model Mech"; date: 
            "2011-07"; volume: "4"; issue: "4"; pages: "537-47"
COMMENT     Sequence Label: pMCS.71/2272.Hygro
FEATURES             Location/Qualifiers
     source          1..5134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..35
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    44..77
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        101..600
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(136..159)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     533..552
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    2129..2162
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     2171..2187
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2171..2187
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2221..2237)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2222..2238)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    2240..2273
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        complement(2292..2310)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2331..2347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2331..2347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2344..2366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2355..2371
                     /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(2379..2385,2386..2403,2404..2409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2424..2445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2562..2579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2733..3321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2813..2832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3492..4283,4284..4352))
                     /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     4115..4134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4353..4457)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4483..4938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4620..4641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4832..4851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5065..5087
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5079..5096
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5080..5096
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5103..5122
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5103..5121
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cataacttcg tataatgtat gctatacgaa cggtagcggc cgcgaagttc ctattctcta
       61 gaaagtatag gaacttcgga tccccccgat gaattctacc gggtagggga ggcgcttttc
      121 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
      181 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
      241 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
      301 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
      361 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
      421 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
      481 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
      541 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
      601 accgatccag ccgccaccat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt
      661 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
      721 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
      781 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt
      841 ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt
      901 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
      961 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc
     1021 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     1081 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     1141 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     1201 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     1261 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     1321 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
     1381 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     1441 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     1501 gcaatcgtcc gatccggatg ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     1561 ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     1621 cactcgtccg agggcaaagg aatagtcgag aaattgatga tctattaaac aataaagatg
     1681 tccactaaaa tggaagtttt tcctgtcata ctttgttaag aagggtgaga acagagtacc
     1741 tacattttga atggaaggat tggagctacg ggggtggggg tggggtggga ttagataaat
     1801 gcctgctctt tactgaaggc tctttactat tgctttatga taatgtttca tagttggata
     1861 tcataattta aacaagcaaa accaaattaa gggccagctc attcctccca ctcatgatct
     1921 atagatctat agatctctcg tgggatcatt gtttttctct tgattcccac tttgtggttc
     1981 taagtactgt ggtttccaaa tgtgtcagtt tcatagcctg aagaacgaga tcagcagcct
     2041 ctgttccaca tacacttcat tctcagtatt gttttgccaa gttctaattc catcagaagc
     2101 tgggctgcag gaattcgata tcaagcttga agttcctatt ctctagaaag tataggaact
     2161 tcgcggccgc tctagaacta gtggatcccc cgggctgcag gaattcgata tcaagcttat
     2221 cgataccgtc gacctcgaga taacttcgta taaagtatcc tatacgaagt tatggtaccc
     2281 agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg
     2341 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2401 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2461 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2521 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2581 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2641 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2701 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2761 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2821 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2881 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2941 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3001 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3061 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3121 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     3181 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3241 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3301 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3361 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3421 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3481 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3541 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3601 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3661 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3721 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3781 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3841 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3901 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3961 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4021 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4081 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4141 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4201 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4261 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4321 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4381 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4441 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     4501 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4561 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4621 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4681 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4741 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4801 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4861 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4921 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     4981 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     5041 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     5101 tgtaatacga ctcactatag ggcgaattgg agct
//
