LOCUS       sequence-465438-        7620 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7620)
  TITLE     Hurley Lab Plasmids 2025
REFERENCE   2  (bases 1 to 7620)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pCAG/TSF-VPS34
FEATURES             Location/Qualifiers
     source          1..7620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(86..105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      899..1487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1388..1407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1641..1658
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    1726..1860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1750..1769)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1804..1823
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(1866..2061)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1880..2015
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    2120..2141
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2156..2161,2162..2179,2180..2186)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2194..2210
                     /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     2199..2221
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2218..2234
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2218..2234
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2577..2596
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2595..2650
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2696..2715
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(5475..5498)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5505..5528)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(5565..5588)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     regulatory      5594..5603
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5650..5669)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5677..6693
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5731..5753)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(6694..6971)
                     /label=chicken beta-actin promoter
     enhancer        6973..7352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7383..7487
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7488..7556,7557..7620,1..728)
                     /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"
ORIGIN
        1 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
       61 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      121 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      181 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      241 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      301 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
      361 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
      421 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
      481 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
      541 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
      601 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      661 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
      721 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
      781 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
      841 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      901 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      961 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1021 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1081 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1141 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1201 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1261 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1321 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1381 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1441 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1501 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     1561 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     1621 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     1681 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcggatcc agacatgata
     1741 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1801 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1861 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     1921 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     1981 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
     2041 gttgctgact aattgagatg cggatccgct ggcacgacag gtttcccgac tggaaagcgg
     2101 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     2161 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     2221 gaaacagcta tgaccatgat tacgccaagc ttgggctgca ggtcgaggga tctccataag
     2281 agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct ggctagtaaa
     2341 acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca aaatataaaa
     2401 aaaatctaac ctcaagtcaa ggcttttcta tggaataagg aatggacagc agggggctgt
     2461 ttcatatact gatgacctct ttatagccaa cctttgttca tggcagccag catatgggca
     2521 tatgttgcca aactctaaac caaatactca ttctgatgtt ttaaatgatt tgccctccca
     2581 tatgtccttc cgagtgagag acacaaaaaa ttccaacaca ctattgcaat gaaaataaat
     2641 ttcctttatt agccagaagt cagatgctca aggggcttca tgatgtcccc ataatttttg
     2701 gcagagggaa aaagatctca gtggtatttg tgagccaggg cattggccac accagccacc
     2761 accttctgat aggcagcctg cacctgagga gtgaattcct cgagtcattt gcgccagtac
     2821 tgggcaaact tgtgaatctg ttccaccaca gcggcgaaca gagcatgcac actctcgtcg
     2881 atcagagact gcatatagtg cacggcttcc tcatcagaca ggtccagcct aaatttatcc
     2941 tgcacctttt tcacggtctt gtcgggctcc agagcaatat ctgggatatt ggcgtccacc
     3001 atcaggctga acagattcag gatcaggttt gagtatctcc gcaggtgcag aaaggcagtg
     3061 taacactgct tgcggaattc ctgatactgc tcggactgtg ttccccccat gccttccacc
     3121 atctccttat tcagtttcat aggtggtggc agtggctttg gatcccttcc cagaatatac
     3181 ccaaagtcga tgtggaacag cttcccggtt ttagtcagca gcaggttatc cagatgcctg
     3241 tcgcccactc ccagaatgta ggtgatcaca caatagccgg cgcagctttt cacgtaagta
     3301 tccatcactt cagcactgat cccattaggg ccgttctcag agggggcata cttcctaaag
     3361 aaattctgaa tgctcccttc agtgtccagc acctcggcca caggcactga ctggataaac
     3421 tgcatgaagc cgtgctttgt actggtagcc agcaccttgt agggtgtcag tttcagatcc
     3481 aggttctcct tgcgcagcag tttgtccatc agagaaatga tctgcagaat cagctgatcc
     3541 tgcctcagat cgtcgccatg tttaaagatc acgggatact tgccgccgtc ttctgtttta
     3601 aagaacagct gagctggcat cagggcggat ttgaacagag tggctgtctc aggaatgatt
     3661 cctctaatct tcacctgggg ttccagaggc agggggatca gctccacatc actcagattc
     3721 attttctcgt tgtctcccag cagggcctgc agtctttcat tctttttctt gcggttccca
     3781 gactccctct gcacagcctt catcaggtgc accagccgat ccacaaaggt ctgctgagcg
     3841 gccagcagag agcgcatcac cctcacggac ttgtcccctt tcagcagggc ctggctgaag
     3901 cgcctcatca cgttcaggta catctcatga gtctttgggt ctctctgctg tgtatcctgg
     3961 tcttcacact ccacaatcac gtaccaatac aggtaattag ccagtgtgct gttcttacag
     4021 gcccgtgaga tcaggaaggt gcacagatcc tgttccagat tctcgccgtc aggcacctcc
     4081 tttgttttgg aggcaggggg tggactagac acgcttggca gaggtgaggt aatgatctgg
     4141 gagctatcaa tttcagcgct gttgattccg ctatttgaca cgttctcgga cactgaactc
     4201 tggctgtctt tcttggtggg ttccagccca tttttgatat cgtcgaagtt ctcgtacttc
     4261 agggcctgca ccagctgcag cagatacatc agcagatcct catcgtcagc ctgcctcagc
     4321 ctagccacag cgtatctccg cactgttgga ttggtatagt gagaggacag cagttccagg
     4381 ctatcctcca cgtccattgg cttccatttg cccagcagtt ccagagcctg cttggcctcc
     4441 tgaggcaggt cccaattcac gcacttcaga aatttggtca gggctttctc ctggttagtc
     4501 agatagtagc ggaacttcca caccagatcc tgttcctcgt aggtcagctg tttagtaggg
     4561 ggatagctca caatgatatt cagctggtcc cttgtagcgg cgtttggctt cagatcatgg
     4621 tcactagggc cagatctcag ggaccgggcc agcttatggt gttttgattc caccaggttc
     4681 tccatactca tctgggggtc tggcactttc accagctcaa agctggtcag aatggggctt
     4741 gattcatcgc cgtccttctc atagtacacg attccgtatt ccttatcgtc acatttcacg
     4801 catctaaact ccaccatcag atacatgaaa ttactagacc gtttttcgct ctcgttaatc
     4861 atttcgatct ctctgaaggt cagccgatcc agccagtcca ccttcaccat atgtccctgg
     4921 cggtgagcct tagtcagttt ggccagcctt gacatctggt cctcactcag agtggagctt
     4981 gttctcccgg gggtcttagt tggttcagag ccatcggcct ccacattggg ccacactttc
     5041 aggtcgtgca tcccctgccg aaacattccg tacttcccga acagggacac tgtggtccct
     5101 cccactggca cagcctttcc aggtccgtac acatcccaga tggtcagagc cacctgggcg
     5161 tttcttggca ggtcaggata cttcacaggc agtttcagcc attcattcca gttccatctt
     5221 gtagaaaagg ctttgtagga ggtcctcaca ggcagagcca gtggctttcc ctcagcgaac
     5281 acctgacaag tcacgtacag atcggagcat gtttcctgat acagtccgct aaacttcagc
     5341 atggggtcct ccagcacggc cttataagat ttctgttccc gtttgccctc cagggatcca
     5401 atcttcagct gcacgttgat atccaggtcg caggaataga tgtaatggaa cttttcagcc
     5461 tcgcccatgg tacccttgtc atcgtcatcc ttgtaatcga atcctttttc gaactgcggg
     5521 tggctccacg atccacctcc cgatccacct cgggcacctc cacctttctc gaactgcggg
     5581 tggctccagc tagccatggt ggcatcgata ccgtcgatcg aggaattctt tgccaaaatg
     5641 atgagacagc acaataacca gcacgttgcc caggagctgt aggaaaaaga agaaggcatg
     5701 aacatggtta gcagaggctc tagagccgcc ggtcacacgc cagaagccga accccgccct
     5761 gccccgtccc ccccgaaggc agccgtcccc ccgcggacag ccccgaggct ggagagggag
     5821 aaggggacgg cggcgcggcg acgcacgaag gccctccccg cccatttcct tcctgccggc
     5881 gccgcaccgc ttcgccccgc gcccgctaga gggggtgcgg cggcgcctcc cagatttcgg
     5941 ctccgcacag atttgggaca aaggaagtcc ctgcgccctc tcgcacgatt accataaaag
     6001 gcaatggctg cggctcgccg cgcctcgaca gccgccggcg ctccgggggc cgccgcgccc
     6061 ctcccccgag ccctccccgg cccgaggcgg ccccgccccg cccggcaccc ccacctgccg
     6121 ccaccccccg cccggcacgg cgagccccgc gccacgcccc gtacggagcc ccgcacccga
     6181 agccgggccg tgctcagcaa ctcggggagg ggggtgcagg gggggttgca gcccgaccga
     6241 cgcgcccaca ccccctgctc acccccccac gcacacaccc cgcacgcagc ctttgttccc
     6301 ctcgcagccc cccccgcacc gcggggcacc gcccccggcc gcgctcccct cgcgcacact
     6361 gcggagcgca caaagccccg cgccgcgccc gcagcgctca cagccgccgg gcagcgcgga
     6421 gccgcacgcg gcgctcccca cgcacacaca cacgcacgca ccccccgagc cgctcccccc
     6481 gcacaaaggg ccctcccgga gcccctcaag gctttcacgc agccacagaa aagaaacaag
     6541 ccgtcattaa accaagcgct aattacagcc cggaggagaa gggccgtccc gcccgctcac
     6601 ctgtgggagt aacgcggtca gtcagagccg gggcgggcgg cgcgaggcgg cggcggagcg
     6661 gggcacgggg cgaaggcagc gcgcagcgac tcccgcccgc cgcgcgcttc gctttttata
     6721 gggccgccgc cgccgccgcc tcgccataaa aggaaacttt cggagcgcgc cgctctgatt
     6781 ggctgccgcc gcacctctcc gcctcgcccc gccccgcccc tcgccccgcc ccgccccgcc
     6841 tggcgcgcgc cccccccccc cccccgcccc catcgctgca caaaataatt aaaaaataaa
     6901 taaatacaaa attgggggtg gggagggggg ggagatgggg agagtgaagc agaacgtggg
     6961 gctcacctcg accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     7021 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     7081 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     7141 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     7201 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     7261 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     7321 ttactattaa taactagtca ataatcaatg tcgacccagg tggcactttt cggggaaatg
     7381 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     7441 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     7501 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7561 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
//
