LOCUS       sequence-465437-        7493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7493)
  TITLE     Hurley Lab Plasmids 2025
REFERENCE   2  (bases 1 to 7493)
  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"
COMMENT     Sequence Label: pCAG/VPS34(R561D|R566D|K567D|K568D)
FEATURES             Location/Qualifiers
     source          1..7493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     137..159
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     221..240
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      288..297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3047..3066)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3112..3167
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3166..3185)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3528..3544)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3528..3544)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3541..3563)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3552..3568
                     /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(3576..3582,3583..3600,3601..3606))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3621..3642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3701..3896
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3747..3882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3902..4036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3939..3958)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3993..4012
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4104..4121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4275..4863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4355..4374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5034..5825,5826..5894))
                     /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     5657..5676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5895..5999)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6030..6409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          join(6690..7493,1..213)
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
ORIGIN
        1 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
       61 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
      121 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
      181 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
      241 ctgtctcatc attttggcaa agaattcctc gatcgacggt atcgatagcc accatgggcg
      301 aggctgaaaa gttccattac atctattcct gcgacctgga tatcaacgtg cagctgaaga
      361 ttggatccct ggagggcaaa cgggaacaga aatcttataa ggccgtgctg gaggacccca
      421 tgctgaagtt tagcggactg tatcaggaaa catgctccga tctgtacgtg acttgtcagg
      481 tgttcgctga gggaaagcca ctggctctgc ctgtgaggac ctcctacaaa gccttttcta
      541 caagatggaa ctggaatgaa tggctgaaac tgcctgtgaa gtatcctgac ctgccaagaa
      601 acgcccaggt ggctctgacc atctgggatg tgtacggacc tggaaaggct gtgccagtgg
      661 gagggaccac agtgtccctg ttcgggaagt acggaatgtt tcggcagggg atgcacgacc
      721 tgaaagtgtg gcccaatgtg gaggccgatg gctctgaacc aactaagacc cccgggagaa
      781 caagctccac tctgagtgag gaccagatgt caaggctggc caaactgact aaggctcacc
      841 gccagggaca tatggtgaag gtggactggc tggatcggct gaccttcaga gagatcgaaa
      901 tgattaacga gagcgaaaaa cggtctagta atttcatgta tctgatggtg gagtttagat
      961 gcgtgaaatg tgacgataag gaatacggaa tcgtgtacta tgagaaggac ggcgatgaat
     1021 caagccccat tctgaccagc tttgagctgg tgaaagtgcc agacccccag atgagtatgg
     1081 agaacctggt ggaatcaaaa caccataagc tggcccggtc cctgagatct ggccctagtg
     1141 accatgatct gaagccaaac gccgctacaa gggaccagct gaatatcatt gtgagctatc
     1201 cccctactaa acagctgacc tacgaggaac aggatctggt gtggaagttc cgctactatc
     1261 tgactaacca ggagaaagcc ctgaccaaat ttctgaagtg cgtgaattgg gacctgcctc
     1321 aggaggccaa gcaggctctg gaactgctgg gcaaatggaa gccaatggac gtggaggata
     1381 gcctggaact gctgtcctct cactatacca atccaacagt gcggagatac gctgtggcta
     1441 ggctgaggca ggctgacgat gaggatctgc tgatgtatct gctgcagctg gtgcaggccc
     1501 tgaagtacga gaacttcgac gatatcaaaa atgggctgga acccaccaag aaagacagcc
     1561 agagttcagt gtccgagaac gtgtcaaata gcggaatcaa cagcgctgaa attgatagct
     1621 cccagatcat tacctcacct ctgccaagcg tgtctagtcc accccctgcc tccaaaacaa
     1681 aggaggtgcc tgacggcgag aatctggaac aggatctgtg caccttcctg atctcacggg
     1741 cctgtaagaa cagcacactg gctaattacc tgtattggta cgtgattgtg gagtgtgaag
     1801 accaggatac acagcagaga gacccaaaga ctcatgagat gtacctgaac gtgatgaggc
     1861 gcttcagcca ggccctgctg aaaggggaca agtccgtgag ggtgatgcgc tctctgctgg
     1921 ccgctcagca gacctttgtg gatcggctgg tgcacctgat gaaggctgtg caggacgagt
     1981 ctgggaacga tgacgataag aatgaaagac tgcaggccct gctgggagac aacgagaaaa
     2041 tgaatctgag tgatgtggag ctgatccccc tgcctctgga accccaggtg aagattagag
     2101 gaatcattcc tgagacagcc actctgttca aatccgccct gatgccagct cagctgttct
     2161 ttaaaacaga agacggcggc aagtatcccg tgatctttaa acatggcgac gatctgaggc
     2221 aggatcagct gattctgcag atcatttctc tgatggacaa actgctgcgc aaggagaacc
     2281 tggatctgaa actgacaccc tacaaggtgc tggctaccag tacaaagcac ggcttcatgc
     2341 agtttatcca gtcagtgcct gtggccgagg tgctggacac tgaagggagc attcagaatt
     2401 tctttaggaa gtatgccccc tctgagaacg gccctaatgg gatcagtgct gaagtgatgg
     2461 atacttacgt gaaaagctgc gccggctatt gtgtgatcac ctacattctg ggagtgggcg
     2521 acaggcatct ggataacctg ctgctgacta aaaccgggaa gctgttccac atcgactttg
     2581 ggtatattct gggaagggat ccaaagccac tgccaccacc tatgaaactg aataaggaga
     2641 tggtggaagg catgggggga acacagtccg agcagtatca ggaattccgc aagcagtgtt
     2701 acactgcctt tctgcacctg cggagatact caaacctgat cctgaatctg ttcagcctga
     2761 tggtggacgc caatatccca gatattgctc tggagcccga caagaccgtg aaaaaggtgc
     2821 aggataaatt taggctggac ctgtctgatg aggaagccgt gcactatatg cagtctctga
     2881 tcgacgagag tgtgcatgct ctgttcgccg ctgtggtgga acagattcac aagtttgccc
     2941 agtactggcg caaatgactc gaggaattca ctcctcaggt gcaggctgcc tatcagaagg
     3001 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     3061 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3121 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     3181 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     3241 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     3301 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     3361 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     3421 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     3481 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     3541 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3601 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3661 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     3721 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3841 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3901 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3961 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4021 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4081 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4141 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4201 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4261 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4441 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4681 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4801 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4861 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4921 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4981 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5041 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5101 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5161 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5221 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5281 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5341 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5401 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5461 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5521 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5581 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5641 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5701 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5761 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5821 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5881 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5941 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6001 acatttcccc gaaaagtgcc acctgggtcg acattgatta ttgactagtt attaatagta
     6061 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6121 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6181 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6241 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6301 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6361 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
     6421 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
     6481 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg gggcggagca
     6541 aggggggggg gggggggggg gggggggggg ggggggcgag gcggagaggt gcggcggcag
     6601 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc
     6661 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc
     6721 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
     6781 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac
     6841 ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt
     6901 gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg
     6961 gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc
     7021 gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg
     7081 aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg
     7141 cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     7201 cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     7261 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     7321 aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt
     7381 gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg
     7441 agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aag
//
