LOCUS       sequence-189851-        6273 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  allows Cre-mediated expression of transgene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6273)
  AUTHORS   Nakamura T, Gulick J, Pratt R, Robbins J
  TITLE     Noonan syndrome is associated with enhanced pERK activity, the 
            repression of which can prevent craniofacial malformations.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Sep 8;106(36):15436-41. doi: 
            10.1073/pnas.0903302106. Epub 2009 Aug 24.
  PUBMED    19706403
REFERENCE   2  (bases 1 to 6273)
  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: 
            "10.1073/pnas.0903302106"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2009-09-8- 8"; volume: "106"; issue: "36"; pages: "15436-41"
COMMENT     Sequence Label: CAG-lox-CAT-lox-unique Bam cloning site-hGH po...
FEATURES             Location/Qualifiers
     source          1..6273
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        119..498
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(530..549)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(531..549)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(559..576)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(559..575)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(568..590)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      717..1172
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(804..823)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1014..1035
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1198..1302
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1303..1371,1372..2163)
                     /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(1521..1540)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2334..2922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2823..2842
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3076..3093
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3210..3231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3246..3251,3252..3269,3270..3276)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3284..3300
                     /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     3289..3311
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3308..3324
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3308..3324
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3343..3363
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3345..3363
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3799..3818
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     protein_bind    4035..4068
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4090..4224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4114..4133)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4168..4187
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4649..4669)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4799..4864
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(5031..5690)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5605..5624
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    5756..5789
                     /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(5840..5859)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(5921..5943)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 cccccccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg
       61 ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
      121 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
      181 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
      241 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
      301 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
      361 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
      421 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
      481 tagtcaataa tcaatgtcga cctcgagggg gggcccggta cccaattcgc cctatagtga
      541 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
      601 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      661 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
      721 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      781 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      841 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      901 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      961 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1021 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1081 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1141 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     1201 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1261 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1321 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1381 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1441 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1501 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1561 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1621 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1681 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1741 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1801 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1861 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1921 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1981 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2041 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2101 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2161 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2221 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2281 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2341 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2401 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2461 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2521 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2581 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2641 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2701 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2761 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2821 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2881 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2941 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3001 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3061 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3121 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3181 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     3241 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     3301 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     3361 agggaacaaa agctggagct ccaccgcggt ggcggccgca acaggcatct actgagtgga
     3421 cccaacgcat gagaggacag tgccaagcaa gcaactcaaa tgtcccaccg gttgggcatg
     3481 gccaggtagc ctatgctgtg tctggacgtc ctcctgctgg tatagttatt ttaaaatcag
     3541 aaggacaggg aagggagcag tggttcacgc ctgtaatccc agcaatttgg gaggccaagg
     3601 tgggtagatc acctgagatt aggagttgga gaccagcctg gccaatatgg tgaaaccccg
     3661 tctctaccaa aaaaacaaaa attagctgag cctggtcatg catgcctgga atcccaacaa
     3721 ctcgggaggc tgaggcagga gaatcgcttg aacccaggag gcggagattg cagtgagcca
     3781 agattgtgcc actgcactcc agcttggttc ccaatagacc ccgcaggccc tacaggttgt
     3841 cttcccaact tgccccttgc tccataccac ccccctccac cccataatat tatagaagga
     3901 cacctagtca gacaaaatga tgcaacttaa ttttattagg acaaggctgg tgggcactgg
     3961 agtggcaact tccagggcca ggagaggcac tggggagggg tcacagggat gccaccggat
     4021 ccggaaccct taatataact tcgtataatg tatgctatac gaagttatta ggtccctcga
     4081 ccggggatcg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     4141 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     4201 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4261 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     4321 gattatgatc tctagtcaag gcactataca tcaaatattc cttattaacc cctttacaaa
     4381 ttaaaaagct aaaggtacac aatttttgag catagttatt aatagcagac actctatgcc
     4441 tgtgtggagt aagaaaaaac agtatgttat gattataact gttatgccta cttataaagg
     4501 ttacagaata tttttccata attttcttgt atagcagtgc agctttttcc tttgtggtgt
     4561 aaatagcaaa gcaagcaaga gttctattac taaacacagc atgactcaaa aaacttagca
     4621 attctgaagg aaagtccttg gggtcttcta cctttctctt cttttttgga ggagtagaat
     4681 gttgagagtc agcagtagcc tcatcatcac tagatggcat ttcttctgag caaaacaggt
     4741 tttcctcatt aaaggcattc caccactgct cccattcatc agttccatag gttggaatct
     4801 aaaatacaca aacaattaga atcagtagtt taacacatta tacacttaaa aattttatat
     4861 ttaccttaga gctttaaatc tctgtaggta gtttgtccaa ttatgtcaca ccacagaagt
     4921 aaggttcctt cacaaagatc cggcgaattt ctgccattca tccgcttatt atcacttatt
     4981 caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg
     5041 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     5101 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     5161 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     5221 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     5281 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     5341 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     5401 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     5461 attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     5521 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     5581 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     5641 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     5701 ttagctcctg aaaatctcgc caagctcctc gagatccaga tccggaaccc ttaatataac
     5761 ttcgtataat gtatgctata cgaagttatt aggtccctcg accgcccagc ttgaattctt
     5821 tgccaaaatg atgagacagc acaataacca gcacgttgcc caggagctgt aggaaaaaga
     5881 agaaggcatg aacatggtta gcagaggctc tagagccgcc ggtcacacgc cagaagccga
     5941 accccgccct gccccgtccc ccccgaaggc agccgtcccc ccgcggacag ccccgaggct
     6001 ggagagggag aaggggacgg cggcgcggcg acgcacgaag gccctccccg cccatttcct
     6061 tcctgccggc gccgcaccgc ttcgccccgc gcccgctaga gggggtgcgg cggcgcctcc
     6121 cagatttcgg ctccgcacag atttgggaca aaggaagtcc ctgcgccctc tcgcacgatt
     6181 accataaaag gcaatggctg cggctcgccg cgcctcgaca gccgccggcg ctccgggggc
     6241 cgccgcgccc ctcccccgag ccctccccgg ccc
//
