LOCUS       sequence-465551-        7786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of EGFP-tagged PODXL WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7786)
  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 7786)
  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: pcDNA3-EGFP/VSVG-rbPodxl
FEATURES             Location/Qualifiers
     source          1..7786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..40)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     267..287
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(296..724)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(406..427)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     618..637
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    770..994
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     983..1000
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        1020..1038
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1020..1037
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(join(2660..3370,3371..3373,3374..3376))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMTDKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2702..2723)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3051..3070
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3310..3331
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3377..3409)
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     primer_bind     complement(3491..3510)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3492..3510)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3555..3758)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3584..3604)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3759..4138
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4310..4329
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4408..4512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4513..4581,4582..5373)
                     /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(4731..4750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5544..6132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6033..6052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6286..6303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6420..6441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6456..6461,6462..6479,6480..6486)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6494..6510
                     /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     6499..6521
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6518..6534
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6518..6534
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6583..6704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6594..6613)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6648..6667
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6878..7672)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6989..7008)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7599..7618
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7739..7786,1..282))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7753..7786,1..102)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
       61 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
      121 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
      181 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
      241 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gaattaattc
      301 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
      361 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
      421 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
      481 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
      541 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
      601 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
      661 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
      721 gcgtggggat accccctaga gccccagctg gttctttccg cctcagaagc catagagccc
      781 accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
      841 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
      901 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
      961 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg agctctagca
     1021 tttaggtgac actatagaat agggccctct agatgcatgc tcgagcggcc gcttagaggt
     1081 gcgtgtcctc ctcctcgtcc aggtcgtcct tggtcaggtt gtccagcggg acaatccagc
     1141 tgtcccccag ctccccgtta aggttgacca ccttcttctc ctgcatctcg gccgacgtct
     1201 ccatcacttc tagggtcggg ttgtcatggt aaccgttctc cacggtctgc agctcctccg
     1261 tgagtcgttg ctggtccttc ctatgggaga ggcgctggtg gcagcagccg tagagggccg
     1321 cgacaagcag caggaaggag gccatgcaga cgatggtgat gatgagcggc aggctgaagc
     1381 ggtcctcggt ctcctccggg ggcccctggt cccccagctg catgtcgctg acgccctcct
     1441 ctctcaggtc atcccatctg tccttcagca gctcgaacac ggccgtgggc agcagattcg
     1501 tcttgatggt gatttctttg accaccaccg cgtggctccc cagcatgggc gccagcagca
     1561 cgtggcactg gtcctgggcg gggttgaagg tgggcttggc cgctcggcac agcagggtga
     1621 ccagtttgtc ttccggaagg ctgtctgtgc aggggctgct tctggtgagg ttcaggatga
     1681 gcagctgctc ggttggtctc tccggaggcc cgcagcttac cctctggtct ctcactgccg
     1741 aggtggggga aagcgtgctg gggccgtgca gggcagttga agccaactca gtgctggagc
     1801 ttgtggggcc cctgccggtt gtgtcctctg ttcctgccgc cgaggggccc gagggctgca
     1861 gtgcggggga gggggaggga gaggagggaa agggggagga ggagggggag gctgggctgg
     1921 gtggcatcgg gctgggcatc ccttgagtct ctgctgagct ggccacgggc gtcagtgtcg
     1981 caggcttccc ctggggtgtg gagagggtgc cagagccggt gccagagcca gtgacagggc
     2041 ccaggctgcc agcagccgtg gggaaactga gctggggccc ctcggagggc ttcggggtgg
     2101 catggcccag caggtcaggg ctggtcgagt tggtcacagg aggagcagtg ggtcccgggg
     2161 tgccgacatc tgtggtctga cggggggcct ggctcgtggc ctctgtggcc agaggctttg
     2221 tggtgatgct gtggttgctc ggagctttgg atgcagctgg gtcctgagct gcgacctggc
     2281 tgccactggt gacaggccga gcagagccgg tgacgtgtgc cgtggggacg ccacctgcat
     2341 ccttagcttc agccttagta gtaacactca ggctctgctg tgttgtggtc gagccctggg
     2401 ccggggctga gttctccacg gagcttccag gggatgcaga ggggctctgg gtggctgggg
     2461 ctttcgtcgt catgggtgta gtgttctgct cggccgggac cgaagccgcc acggagcttt
     2521 tgggggctgg ggctgaggct ggggcaggcc tggtagatgt tgaagtggcc atgggggttg
     2581 ggccgggctg gggactcttt tcctggaatt ccgtcgcatt ggagaagggc acgtagaagt
     2641 tagggccttc tgtgccattc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     2701 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
     2761 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     2821 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     2881 agttcacctt gatgccgttc ttctgcttgt cggtcatgat atagacgttg tggctgttgt
     2941 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     3001 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     3061 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     3121 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     3181 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     3241 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     3301 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     3361 cgcccttgct caccatctta cccaatcgat tcatttcgat atcagtatac ggcctggcgg
     3421 catgcagcag caaggctagc ggcaggagca aggcggtcac tggtaaggcc atggtaccaa
     3481 gcttgggtct ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct
     3541 ctagttagcc agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     3601 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
     3661 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     3721 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     3781 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     3841 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     3901 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     3961 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     4021 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     4081 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa
     4141 cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct aagcagattc
     4201 ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac
     4261 tcagcgacct ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag
     4321 cagattgtac tgagagtgca ccatagggga tcgggagatc gatctcccga tccgtcgacg
     4381 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4441 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4501 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4561 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4621 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4681 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4741 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4801 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4861 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4921 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4981 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5041 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5101 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5161 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5221 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5281 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5341 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5401 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5461 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5521 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5581 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5641 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5701 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5761 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5821 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5881 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5941 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6001 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6061 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6181 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6241 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6301 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6361 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6421 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6481 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6541 tacgccaagc tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt
     6601 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     6661 ctattgcttt atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac
     6721 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     6781 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     6841 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     6901 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     6961 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     7021 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     7081 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     7141 tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     7201 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     7261 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     7321 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     7381 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     7441 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat
     7501 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     7561 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     7621 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     7681 atcctcatcc tgtctcttga tcagatccga aaatggatat acaagctccc gggagctttt
     7741 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaa
//
