LOCUS       sequence-225209-        3006 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 3006)
  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 3006)
  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
FEATURES             Location/Qualifiers
     source          1..3006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(13..31)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(52..68)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(52..68)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(65..87)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    76..92
                     /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(100..106,107..124,125..130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    145..166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(283..300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(454..1042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(534..553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1213..2004,2005..2073))
                     /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     1836..1855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2074..2178)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2204..2659)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2341..2362)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2553..2572
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2786..2808
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2800..2817
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2801..2817
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2824..2843
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2824..2842
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2857..2890
                     /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)."
     primer_bind     2899..2915
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2899..2915
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2949..2965)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2950..2966)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    2968..3001
                     /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)."
ORIGIN
        1 cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct
       61 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
      121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
      181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      241 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
      301 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      361 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      421 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      481 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      541 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      601 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      661 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      721 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      781 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      841 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      901 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      961 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1021 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1081 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1141 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1201 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1261 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1321 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1381 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1441 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1501 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1561 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1621 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1681 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1741 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1801 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1861 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1921 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1981 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2041 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2101 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2161 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     2221 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2281 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2341 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2401 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2461 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2521 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2581 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2641 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2701 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2761 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     2821 ttgtaatacg actcactata gggcgaattg gagctcataa cttcgtataa tgtatgctat
     2881 acgaacggta gcggccgctc tagaactagt ggatcccccg ggctgcagga attcgatatc
     2941 aagcttatcg ataccgtcga cctcgagata acttcgtata aagtatccta tacgaagtta
     3001 tggtac
//
