LOCUS       sequence-465278-        7508 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 7508)
  TITLE     Hurley Lab Plasmids 2025
REFERENCE   2  (bases 1 to 7508)
  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(W885D/Y884D)
FEATURES             Location/Qualifiers
     source          1..7508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(776..795)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    841..896
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(895..914)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1257..1273)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1257..1273)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1270..1292)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1281..1297
                     /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(1305..1311,1312..1329,1330..1335))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1350..1371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1430..1625
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1476..1611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1631..1765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1668..1687)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1722..1741
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1833..1850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2004..2592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2084..2103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2763..3554,3555..3623))
                     /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     3386..3405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3624..3728)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3759..4138
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          4434..5450
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5374..5396
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5458..5477
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      5525..5534
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aggagaacct ggatctgaaa ctgacaccct acaaggtgct ggctaccagt acaaagcacg
       61 gcttcatgca gtttatccag tcagtgcctg tggccgaggt gctggacact gaagggagca
      121 ttcagaattt ctttaggaag tatgccccct ctgagaacgg ccctaatggg atcagtgctg
      181 aagtgatgga tacttacgtg aaaagctgcg ccggctattg tgtgatcacc tacattctgg
      241 gagtgggcga caggcatctg gataacctgc tgctgactaa aaccgggaag ctgttccaca
      301 tcgactttgg gtatattctg ggaagggatc caaagccact gccaccacct atgaaactga
      361 ataaggagat ggtggaaggc atggggggaa cacagtccga gcagtatcag gaattccgca
      421 agcagtgtta cactgccttt ctgcacctgc ggagatactc aaacctgatc ctgaatctgt
      481 tcagcctgat ggtggacgcc aatatcccag atattgctct ggagcccgac aagaccgtga
      541 aaaaggtgca ggataaattt aggctggacc tgtctgatga ggaagccgtg cactatatgc
      601 agtctctgat cgacgagagt gtgcatgctc tgttcgccgc tgtggtggaa cagattcaca
      661 agtttgccca ggacgatcgc aaatgactcg aggaattcac tcctcaggtg caggctgcct
      721 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
      781 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
      841 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
      901 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
      961 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     1021 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     1081 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     1141 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     1201 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     1261 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1321 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1381 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     1441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     1501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     1561 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     1621 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1681 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1741 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     1801 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1861 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1921 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1981 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2041 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2101 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2161 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2221 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2281 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2341 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2401 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2461 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2521 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2581 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2641 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2701 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2761 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2821 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2881 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2941 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3001 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3061 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3121 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3181 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3241 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3301 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3361 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3421 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3481 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3541 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3601 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3661 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3721 gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtta
     3781 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     3841 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3901 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3961 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     4021 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     4081 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     4141 cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat
     4201 tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg
     4261 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     4321 cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
     4381 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
     4441 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
     4501 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
     4561 gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg
     4621 aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg
     4681 tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc
     4741 gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg
     4801 ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     4861 gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc
     4921 cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg
     4981 ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc
     5041 ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc
     5101 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     5161 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta
     5221 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     5281 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg
     5341 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     5401 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     5461 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgat cgacggtatc
     5521 gatagccacc atgggcgagg ctgaaaagtt ccattacatc tattcctgcg acctggatat
     5581 caacgtgcag ctgaagattg gatccctgga gggcaaacgg gaacagaaat cttataaggc
     5641 cgtgctggag gaccccatgc tgaagtttag cggactgtat caggaaacat gctccgatct
     5701 gtacgtgact tgtcaggtgt tcgctgaggg aaagccactg gctctgcctg tgaggacctc
     5761 ctacaaagcc ttttctacaa gatggaactg gaatgaatgg ctgaaactgc ctgtgaagta
     5821 tcctgacctg ccaagaaacg cccaggtggc tctgaccatc tgggatgtgt acggacctgg
     5881 aaaggctgtg ccagtgggag ggaccacagt gtccctgttc gggaagtacg gaatgtttcg
     5941 gcaggggatg cacgacctga aagtgtggcc caatgtggag gccgatggct ctgaaccaac
     6001 taagaccccc gggagaacaa gctccactct gagtgaggac cagatgtcaa ggctggccaa
     6061 actgactaag gctcaccgcc agggacatat ggtgaaggtg gactggctgg atcggctgac
     6121 cttcagagag atcgaaatga ttaacgagag cgaaaaacgg tctagtaatt tcatgtatct
     6181 gatggtggag tttagatgcg tgaaatgtga cgataaggaa tacggaatcg tgtactatga
     6241 gaaggacggc gatgaatcaa gccccattct gaccagcttt gagctggtga aagtgccaga
     6301 cccccagatg agtatggaga acctggtgga atcaaaacac cataagctgg cccggtccct
     6361 gagatctggc cctagtgacc atgatctgaa gccaaacgcc gctacaaggg accagctgaa
     6421 tatcattgtg agctatcccc ctactaaaca gctgacctac gaggaacagg atctggtgtg
     6481 gaagttccgc tactatctga ctaaccagga gaaagccctg accaaatttc tgaagtgcgt
     6541 gaattgggac ctgcctcagg aggccaagca ggctctggaa ctgctgggca aatggaagcc
     6601 aatggacgtg gaggatagcc tggaactgct gtcctctcac tataccaatc caacagtgcg
     6661 gagatacgct gtggctaggc tgaggcaggc tgacgatgag gatctgctga tgtatctgct
     6721 gcagctggtg caggccctga agtacgagaa cttcgacgat atcaaaaatg ggctggaacc
     6781 caccaagaaa gacagccaga gttcagtgtc cgagaacgtg tcaaatagcg gaatcaacag
     6841 cgctgaaatt gatagctccc agatcattac ctcacctctg ccaagcgtgt ctagtccacc
     6901 ccctgcctcc aaaacaaagg aggtgcctga cggcgagaat ctggaacagg atctgtgcac
     6961 cttcctgatc tcacgggcct gtaagaacag cacactggct aattacctgt attggtacgt
     7021 gattgtggag tgtgaagacc aggatacaca gcagagagac ccaaagactc atgagatgta
     7081 cctgaacgtg atgaggcgct tcagccaggc cctgctgaaa ggggacaagt ccgtgagggt
     7141 gatgcgctct ctgctggccg ctcagcagac ctttgtggat cggctggtgc acctgatgaa
     7201 ggctgtgcag agggagtctg ggaaccgcaa gaaaaagaat gaaagactgc aggccctgct
     7261 gggagacaac gagaaaatga atctgagtga tgtggagctg atccccctgc ctctggaacc
     7321 ccaggtgaag attagaggaa tcattcctga gacagccact ctgttcaaat ccgccctgat
     7381 gccagctcag ctgttcttta aaacagaaga cggcggcaag tatcccgtga tctttaaaca
     7441 tggcgacgat ctgaggcagg atcagctgat tctgcagatc atttctctga tggacaaact
     7501 gctgcgca
//
