LOCUS       sequence-10393-e        4064 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 4064)
  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 4064)
  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.DT-A
FEATURES             Location/Qualifiers
     source          1..4064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             304..876
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     intron          953..1018
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     primer_bind     complement(1121..1137)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1121..1137)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1169..1188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1170..1188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1195..1212)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1195..1211)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1204..1226)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1353..1808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1440..1459)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1650..1671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1883..1987
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1988..2056,2057..2848)
                     /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     complement(2206..2225)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3019..3607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3508..3527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3761..3778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3895..3916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3931..3936,3937..3954,3955..3961)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3969..3985
                     /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)."
     primer_bind     3974..3996
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3993..4009
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3993..4009
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4030..4048
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cgggcccccc ctcgagcagt gtggttttca agaggaagca aaaagcctct ccacccaggc
       61 ctggaatgtt tccacccaat gtcgagcagt gtggttttgc aagaggaagc aaaaagcctc
      121 tccacccagg cctggaatgt ttccacccaa tgtcgagcaa accccgccca gcgtcttgtc
      181 attggcgaat tcgaacacgc agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca
      241 tattaaggtg acgcgtgtgg cctcgaacac cgagcgaccc tgcaggtcct cgccatggat
      301 cctgatgatg ttgttgattc ttctaaatct tttgtgatgg aaaacttttc ttcgtaccac
      361 gggactaaac ctggttatgt agattccatt caaaaaggta tacaaaagcc aaaatctggt
      421 acacaaggaa attatgacga tgattggaaa gggttttata gtaccgacaa taaatacgac
      481 gctgcgggat actctgtaga taatgaaaac ccgctctctg gaaaagctgg aggcgtggtc
      541 aaagtgacgt atccaggact gacgaaggtt ctcgcactaa aagtggataa tgccgaaact
      601 attaagaaag agttaggttt aagtctcact gaaccgttga tggagcaagt cggaacggaa
      661 gagtttatca aaaggttcgg tgatggtgct tcgcgtgtag tgctcagcct tcccttcgct
      721 gaggggagtt ctagcgttga atatattaat aactgggaac aggcgaaagc gttaagcgta
      781 gaacttgaga ttaattttga aacccgtgga aaacgtggcc aagatgcgat gtatgagtat
      841 atggctcaag cctgtgcagg aaatcgtgtc aggcgatctc tttgtgaagg aaccttactt
      901 ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat
      961 aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat
     1021 tccaacctat ggaactgatg aatgggagca gtggtggaat gcagatccac tagagtcgac
     1081 ggtatcgata agcttgatat cgaattcctg cagcccgggg gatccactag ttctagagcg
     1141 gccgccaccg cggtggagct ccaattcgcc ctatagtgag tcgtattaca attcactggc
     1201 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     1261 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     1321 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     1381 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1441 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1501 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1561 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1621 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1681 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     1741 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     1801 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttg
     1861 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1921 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1981 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2041 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2101 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2161 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2221 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2281 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2341 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2401 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2461 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2521 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2581 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2641 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2701 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2761 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2821 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2881 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2941 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3001 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3061 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3121 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3781 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3841 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     3901 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     3961 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     4021 caagctcgaa attaaccctc actaaaggga acaaaagctg gtac
//
