LOCUS       sequence-346240-        8455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For lentiviral transduction of 3FLAG-RAB11A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8455)
  AUTHORS   Park H, Hundley FV, Yu Q, Overmyer KA, Brademan DR, Serrano L, Paulo
            JA, Paoli JC, Swarup S, Coon JJ, Gygi SP, Wade Harper J
  TITLE     Spatial snapshots of amyloid precursor protein intramembrane 
            processing via early endosome proteomics.
  JOURNAL   Nat Commun. 2022 Oct 16;13(1):6112. doi: 10.1038/s41467-022-33881-x.
  PUBMED    36245040
REFERENCE   2  (bases 1 to 8455)
  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: "Nat 
            Commun."; date: "2022-10-16"; volume: "13(1):6112. doi"; pages: " 
            10.1038/s41467-022-33881-x"
COMMENT     Sequence Label: pHAGE-3FLAG-RAB11A
FEATURES             Location/Qualifiers
     source          1..8455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     355..372
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    612..633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(648..653,654..671,672..678)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    686..702
                     /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     691..713
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     710..726
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     710..726
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     776..795
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             987..1620
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1667..1792
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2289..2522
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2707..2751
                     /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    3048..3165
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3217..3520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3521..3724
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3675..3695
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3721..3745
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3821..3830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3830..3853
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3857..3880
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3884..3907
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3923..3947
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4596..4620)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4633..5201
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4651..5203
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4795..4812)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5022..5041
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5215..5814
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5215..5234)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5711..5731
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5831..6419
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5884..5904)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6302..6313)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6494..6727
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6846..6864)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6932..7036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7037..7105,7106..7897)
                     /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(7255..7274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8068..8455,1..201)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
       61 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      121 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      181 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      241 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
      301 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
      361 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
      421 ccgattcatt aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg
      481 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
      541 gcttttgcaa aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag
      601 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
      661 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
      721 tatgacatga ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
      781 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
      841 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
      901 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
      961 taaatatgta ctacaaactt agtagttgga agggctaatt cactcccaaa gaagacaaga
     1021 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac
     1081 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     1141 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt
     1201 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg
     1261 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata
     1321 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg
     1381 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg
     1441 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     1501 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     1561 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     1621 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     1681 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     1741 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     1801 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     1861 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     1921 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     1981 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     2041 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     2101 aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga
     2161 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     2221 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     2281 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     2341 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     2401 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     2461 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     2521 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     2581 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     2641 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     2701 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     2761 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     2821 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2881 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2941 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     3001 cgacggtatc gccgaattaa ttcacaaatg gcagtattca tccacaattt taaaagaaaa
     3061 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     3121 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     3181 gacagcagag atccagtttg gactagtgga gttccgcgtt acataactta cggtaaatgg
     3241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     3421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     3481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     3601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     3661 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     3721 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
     3781 tagaagacac cggcggccgc agatctctcg aggttgatct gccaccatgg actacaagga
     3841 tgacgatgac aagggcgatt acaaggatga cgacgataag ggagattaca aggatgacga
     3901 cgataaggct cgaggttatc aaacaagttt gtacaaaaaa gttggcatgg gcacccgcga
     3961 cgacgagtac gactacctct ttaaagttgt ccttattgga gattctggtg ttggaaagag
     4021 taatctcctg tctcgattta ctcgaaatga gtttaatctg gaaagcaaga gcaccattgg
     4081 agtagagttt gcaacaagaa gcatccaggt tgatggaaaa acaataaagg cacagatatg
     4141 ggacacagca gggcaagagc gatatcgagc tataacatca gcatattatc gtggagctgt
     4201 aggtgcctta ttggtttatg acattgctaa acatctcaca tatgaaaatg tagagcgatg
     4261 gctgaaagaa ctgagagatc atgctgatag taacattgtt atcatgcttg tgggcaataa
     4321 gagtgatcta cgtcatctca gggcagttcc tacagatgaa gcaagagctt ttgcagaaaa
     4381 gaatggtttg tcattcattg aaacttcggc cctagactct acaaatgtag aagctgcttt
     4441 tcagacaatt ttaacagaga tttaccgcat tgtttctcag aagcaaatgt cagacagacg
     4501 cgaaaatgac atgtctccaa gcaacaatgt ggttcctatt catgttccac caaccactga
     4561 aaacaagcca aaggtgcagt gctgtcagaa catctaacca actttcttgt acaaagtggt
     4621 tcgataacga attccgcccc ccccccccta acgttactgg ccgaagccgc ttggaataag
     4681 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     4741 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     4801 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     4861 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     4921 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     4981 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     5041 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     5101 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     5161 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatgacc
     5221 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     5281 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     5341 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5401 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5461 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5521 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     5581 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     5641 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     5701 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     5761 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatgcatc
     5821 gatggatcct aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5881 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5941 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     6001 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     6061 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     6121 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     6181 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     6241 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6301 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     6361 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     6421 tgagatcctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6481 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     6541 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6601 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6661 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6721 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     6781 atatcagaga gtgagaggcc cgggttaatt aaggaaaggg ctagatcatt cttgaagacg
     6841 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     6901 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6961 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7021 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7081 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     7141 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7201 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7261 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7321 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7381 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7441 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7501 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7561 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7621 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7681 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7741 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7801 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7861 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     7921 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7981 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8041 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     8101 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     8161 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     8221 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     8281 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     8341 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8401 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgt
//
