LOCUS       sequence-152509-        8635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pHAGE lentiviral vector with dsRed driven by the human E-cadherin 
            promoter. When used in combination with FUGW-d2GFP-ZEB1 3' UTR, 
            comprises the Z-cad dual sensor. Use 2nd generation packaging 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8635)
  AUTHORS   Toneff MJ, Sreekumar A, Tinnirello A, Hollander PD, Habib S, Li S, 
            Ellis MJ, Xin L, Mani SA, Rosen JM
  TITLE     The Z-cad dual fluorescent sensor detects dynamic changes between 
            the epithelial and mesenchymal cellular states.
  JOURNAL   BMC Biol. 2016 Jun 17;14(1):47.
  PUBMED    27317311
REFERENCE   2  (bases 1 to 8635)
  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"; journalName: "BMC Biol.";
            date: "2016-06-17"; volume: "14(1)"; pages: "47"
COMMENT     Sequence Label: pHAGE-E-cadherin-RFP
FEATURES             Location/Qualifiers
     source          1..8635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    24..149
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    646..879
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1064..1108
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1399..1516
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      2929..2938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2935..3612
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(3065..3083)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3115..3135)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3333..3352
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3527..3550
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    3620..4207
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3672..3692)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4090..4101)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5085..5265
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5381..5399)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5467..5571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5572..5640,5641..6432)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5790..5809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6603..7191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7092..7111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7345..7362
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7602..7623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7638..7643,7644..7661,7662..7668)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7676..7692
                     /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     7681..7703
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7700..7716
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7700..7716
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7766..7785
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8354..8611
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
       61 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      121 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      181 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      241 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      301 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      361 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      421 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      481 agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
      541 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
      601 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
      661 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
      721 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
      781 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
      841 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
      901 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
      961 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     1021 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     1081 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     1141 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagttttgct
     1201 gtacttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     1261 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     1321 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgccga attcacaaat
     1381 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     1441 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     1501 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggactagtag
     1561 acatttctga tcattattcc cattaggagg gtggagaaac tgaggctttg ggaggtggtc
     1621 ctgacctagg gaatcaattt gctgactcac taacccatga agctctacag ttaaaaaaga
     1681 ctagattaaa aaatgagaac tcagtaaagg ggctgaggca ggaggatcgc ctgagttcag
     1741 aaatttgaga tcagcctcgg caacatagtg agatcccctc tctagaaaaa ttttttaaaa
     1801 aattaggccg ctcgaggcag agtgcagtgg ctcacgcctg taatccaaca cttcaggagg
     1861 ctgaagaggg tggatcacct gaggtcagga gttcaagacc agcctggcca acatggtgaa
     1921 accccgtctg tactaaaaat acaaaattag ccggtgtggt ggcacacgcc tgtagtccca
     1981 gctactcaat aggctgagac aggagagtct cttgaacccg gcaggcggag gttgcagtga
     2041 gccgagatcg tgccactgca ctccagcctg ggcaagacag agcgagactc cgtctcaaaa
     2101 aatacaaaca aaacaaacaa acaaaaaatt aggctgctag ctcagtggct catggctcac
     2161 acctgaaatc ctagcacttt gggaggccaa ggcaggagga tcgcttcagc ccaggagttc
     2221 gagaccaggc tgggcaatac agggagacac agcgccccca ctgcccctgt ccgccccgac
     2281 ttgtctctct acaaaaaggc aaaagaaaaa aaaaattagc ctggcgtggt ggtgtgcacc
     2341 tgtactccca gctactagag aggctggggc cagaggaccg cttgagccca ggagttcgag
     2401 gctgcagtga gctgtgatcg caccactgca ctccagcttg ggtgaaagag tgagccccat
     2461 ctccaaaacg aacaaacaaa aaatcccaaa aaacaaaaga actcagccaa gtgtaaaagc
     2521 cctttctgat cccaggtctt agtgagccac cggcggggct gggattcgaa cccagtggaa
     2581 tcagaaccgt gcaggtccca taacccacct agaccctagc aactccaggc tagagggtca
     2641 acgcgtctat gcgaggccgg gtgggcgggc cgtcagctcc gccctgggga ggggtccgcg
     2701 ctgctgattg gctgtggccg gcaggtgaac cctcagccaa tcagcggtac ggggggcggt
     2761 gcctccgggg ctcacctggc tgcagccacg caccccctct cagtggcgtc ggaactgcaa
     2821 agcacctgtg agcttgcgga agtcagttca gactccagcc cgctccagcc cggcccgacc
     2881 cgaccgcacc cggcgcctgc cctcgctcgg cgtccccggc cagccgcggc cgccatggcc
     2941 tcctccgagg acgtcatcaa ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg
     3001 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     3061 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     3121 ccccagttcc agtacggctc caaggtgtac gtgaagcacc ccgccgacat ccccgactac
     3181 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3241 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gctccttcat ctacaaggtg
     3301 aagttcatcg gcgtgaactt cccctccgac ggccccgtaa tgcagaagaa gactatgggc
     3361 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
     3421 aaggccctga agctgaagga cggcggccac tacctggtgg agttcaagtc catctacatg
     3481 gccaagaagc ccgtgcagct gcccggctac tactacgtgg actccaagct ggacatcacc
     3541 tcccacaacg aggactacac catcgtggag cagtacgagc gcgccgaggg ccgccaccac
     3601 ctgttcctgt agggatccta atcaacctct ggattacaaa atttgtgaag attgactggt
     3661 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3721 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3781 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3841 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3901 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3961 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     4021 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     4081 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4141 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4201 tccccgcctg ggatctactc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     4261 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg agggtgggtt
     4321 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     4381 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     4441 atcacctgca gggccggagc tagcggggag ctcacgggga cagccccccc ccaaagcccc
     4501 cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc cggggctccg
     4561 ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac agcccgggca
     4621 cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc
     4681 agacacctgg gggatacggg gaaaatctag atccggagct agcggggagc tcacggggac
     4741 agcccccccc caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag
     4801 cgagccgccc ggggctccgc tccggtccgg cgctcccccc gcatccccga gccggcagcg
     4861 tgcggggaca tgcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc
     4921 tcgctgctct ttgagcctgc agacacctgg gggatacggg gaaaatctag atccggaccc
     4981 agggatgtaa ttacgtccct cccccgctag ggggcagcac ccagggatgt aattacgtcc
     5041 ctcccccgct agggggcagc atccggctgc tttttgcttg tactgggtct ctctggttag
     5101 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5161 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5221 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     5281 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagagggggt
     5341 taattaagga aagggctaga tcattcttga agacgaaagg gcctcgtgat acgcctattt
     5401 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5461 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5521 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5581 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5641 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5701 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5761 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     5821 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5881 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5941 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6001 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6061 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6721 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7261 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7321 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7381 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa gctcatggct
     7441 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     7501 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc gtggcacgac
     7561 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7621 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7681 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt cacaaataaa
     7741 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7801 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
     7861 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
     7921 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttgga
     7981 aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc taccacacac
     8041 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
     8101 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
     8161 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     8221 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
     8281 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
     8341 gggactttcc agggaggcgt ggcctggcgg gactggggag tggcgagccc tcagatcctg
     8401 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     8461 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     8521 tgcttcagta gtgtgtgccc gtcttgttgt gtgactctgg taactagaga tccctcagac
     8581 ccttttagtc aggtgtggaa aatctctagc agtggcgccc gaaccaggga cttga
//
