LOCUS       sequence-389337-        7048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector containing GFP tagged PKCBII..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7048)
  AUTHORS   Bryant DM, Roignot J, Datta A, Overeem AW, Kim M, Yu W, Peng X, 
            Eastburn DJ, Ewald AJ, Werb Z, Mostov KE
  TITLE     A molecular switch for the orientation of epithelial cell 
            polarization.
  JOURNAL   Dev Cell. 2014 Oct 27;31(2):171-87. doi: 
            10.1016/j.devcel.2014.08.027. Epub 2014 Oct 9.
  PUBMED    25307480
REFERENCE   2  (bases 1 to 7048)
  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: 
            "10.1016/j.devcel.2014.08.027"; journalName: "Dev Cell"; date: 
            "2014-10-27- 27"; volume: "31"; issue: "2"; pages: "171-87"
COMMENT     Sequence Label: GFP-PKCBII
FEATURES             Location/Qualifiers
     source          1..7048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        132..435
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        436..639
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     590..610
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      681..690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(687..689,690..692,693..1403)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(687..689,690..692,693..1403)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(732..753)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(993..1012)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1340..1361
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3449..3465)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3450..3466)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3503..3522)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3504..3522)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3529..3546)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3529..3545)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3538..3560)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    3907..4028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3944..3963)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3998..4017
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4035..4490)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4172..4193)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4384..4403
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4517..4621
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4623..4980
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4623..4814
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4646..4666)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4831..4966
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5015..5809
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5015..5809
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5069..5088)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5679..5698
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5997..6016)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6041..6088
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6417..7005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6906..6925
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
       61 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
      121 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      181 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      241 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      301 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      361 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      421 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      481 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      541 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      601 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      661 cgctagtgga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
      721 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      781 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      841 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      901 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      961 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1021 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1081 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1141 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1201 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1261 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1321 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1381 tcggcatgga cgagctgtac aaggcgcgca agatggctga cccggctgcg gggccgccgc
     1441 cgagcgaggg cgaggagagc accgtgcgct tcgcccgcaa aggcgccctc cggcagaaga
     1501 acgtgcatga ggtcaagaac cacaaattca ccgcccgctt cttcaagcag cccaccttct
     1561 gcagccactg caccgacttc atctggggct tcgggaagca gggattccag tgccaagttt
     1621 gctgctttgt ggtgcacaag cggtgccatg aatttgtcac attctcctgc cctggcgctg
     1681 acaagggtcc agcctccgat gacccccgca gcaaacacaa gtttaagatc cacacgtact
     1741 ccagccccac gttttgtgac cactgtgggt cactgctgta tggactcatc caccagggga
     1801 tgaaatgtga cacctgcatg atgaatgtgc acaagcgctg cgtgatgaat gttcccagcc
     1861 tgtgtggcac ggaccacacg gagcgccgcg gccgcatcta catccaggcc cacatcgaca
     1921 gggacgtcct cattgtcctc gtaagagatg ctaaaaacct tgtacctatg gaccccaatg
     1981 gcctgtcaga tccctacgta aaactgaaac tgattcccga tcccaaaagt gagagcaaac
     2041 agaagaccaa aaccatcaaa tgctccctca accctgagtg gaatgagaca tttagatttc
     2101 agctgaaaga atcggacaaa gacagaagac tgtcagtaga gatttgggat tgggatttga
     2161 ccagcaggaa tgacttcatg ggatctttgt cctttgggat ttctgaactt cagaaagcca
     2221 gtgttgatgg ctggtttaag ttactgagcc aggaggaagg cgagtacttc aatgtgcctg
     2281 tgccaccaga aggaagtgag gccaatgaag aactgcggca gaaatttgag agggccaaga
     2341 tcagtcaggg aaccaaggtc ccggaagaaa agacgaccaa cactgtctcc aaatttgaca
     2401 acaatggcaa cagagaccgg atgaaactga ccgattttaa cttcctaatg gtgctgggga
     2461 aaggcagctt tggcaaggtc atgctttcag aacgaaaagg cacagatgag ctctatgctg
     2521 tgaagatcct gaagaaggac gttgtgatcc aagatgatga cgtggagtgc actatggtgg
     2581 agaagcgggt gttggccctg cccgggaagc cgcccttcct gacccagctc cactcctgct
     2641 tccagaccat ggaccgcctg tactttgtga tggagtacgt gaatgggggc gacctcatgt
     2701 atcacatcca gcaagtcggc cggttcaagg agccccatgc tgtattttac gctgcagaaa
     2761 ttgccatcgg tctgttcttc ttacagagta agggcatcat ttaccgtgac ctaaaacttg
     2821 acaacgtgat gctcgattct gagggacaca tcaagattgc cgattttggc atgtgtaagg
     2881 aaaacatctg ggatggggtg acaaccaaga cattctgtgg cactccagac tacatcgccc
     2941 ccgagataat tgcttatcag ccctatggga agtccgtgga ttggtgggca tttggagtcc
     3001 tgctgtatga aatgttggct gggcaggcac cctttgaagg ggaggatgaa gatgaactct
     3061 tccaatccat catggaacac aacgtagcct atcccaagtc tatgtccaag gaagctgtgg
     3121 ccatctgcaa agggctaatg accaaacacc caggcaaacg tctgggttgt ggacctgaag
     3181 gcgaacgtga tatcaaagag catgcatttt tccggtatat tgattgggag aaacttgaac
     3241 gcaaagagat ccagccccct tataagccaa aagcttgtgg gcgaaatgct gaaaacttcg
     3301 accgattttt cacccgccat ccaccagtcc taacacctcc cgaccaggaa gtcatcagga
     3361 atattgacca atcagaattc gaaggatttt cctttgttaa ctctgaattt ttaaaacccg
     3421 aagtcaagag ctaagtagat gtgtagatcg ataccgtcga cctcgagggg gggcccggta
     3481 ccaggtaagt gtacccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
     3541 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     3601 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3661 gttgcgcagc ctgaatggcg aatggagatc caatttttaa gtgtataatg tgttaaacta
     3721 ctgattctaa ttgtttgtgt attttagatt cacagtccca aggctcattt caggcccctc
     3781 agtcctcaca gtctgttcat gatcataatc agccatacca catttgtaga ggttttactt
     3841 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3901 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3961 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4021 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4081 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4141 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4201 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4261 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4321 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4381 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4441 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4501 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4561 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4621 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4681 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4741 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4801 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4861 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4921 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4981 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5041 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5101 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5161 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     5221 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5281 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5341 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5401 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5461 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5521 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5581 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5641 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5701 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5761 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5821 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5881 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5941 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     6001 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     6061 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     6121 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     6181 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6241 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6301 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6361 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6421 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6481 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6541 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6601 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6661 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6721 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6781 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6841 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6901 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6961 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7021 gcctttttac ggttcctggc cttttgct
//
