LOCUS       sequence-420102-        8166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian overexpression vector for C-terminally Myc and Flag and 
            His tagged human Pchd21 (WT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8166)
  AUTHORS   Bell TA, Luce BE, Hakim P, Ananda VY, Dardari H, Nguyen TH, 
            Monshizadeh A, Chao LH
  TITLE     Prominin 1 and Tweety Homology 1 both induce extracellular vesicle 
            formation.
  JOURNAL   Elife. 2024 Aug 13;13:e100061. doi: 10.7554/eLife.100061.
  PUBMED    39168849
REFERENCE   2  (bases 1 to 8166)
  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: "Elife."; 
            date: "2024-08-13"; volume: "13:e100061. doi"; pages: " 
            10.7554/eLife.100061"
COMMENT     Sequence Label: pCDNA3.1-HsPcdh21-Myc-Flag-His
FEATURES             Location/Qualifiers
     source          1..8166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3515..3535
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3554..3583
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3599..3664
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3680..3709
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(3757..3774)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3763..3987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4033..4461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4120..4139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4330..4351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4470..4490)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4475..4804
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4655..4790
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4717..4736
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4871..5665
                     /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(4925..4944)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5535..5554
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5839..5960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5876..5895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5930..5949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6009..6025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6009..6025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6022..6044)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6033..6049
                     /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)."
     promoter        complement(join(6057..6063,6064..6081,6082..6087))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6102..6123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6240..6257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6411..6999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6491..6510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7170..7961,7962..8030))
                     /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     7793..7812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8031..8135)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccatgcggcg gtgcaggtgg gcagcattag ccctgggtct
      961 tctacgattg tgtttggccc aagctaattt tgctccgcac tttttcgata acggggttgg
     1021 gagtacaaat ggtaatatgg ccctgttcag cttacctgag gatacgccgg ttggttctca
     1081 cgtttatacg ttaaacggta ctgatcctga gggtgatcca atatcctatc atatttcgtt
     1141 tgatccgtcc acgcggtcag tcttttcggt cgaccccaca tttggtaaca tcactctcgt
     1201 tgaggaacta gaccgggagc gtgaggacga gatcgaagcg ataatatcca tctctgatgg
     1261 attaaatttg gtagcagaaa aggtggttat tctggtaacc gacgctaatg acgaggctcc
     1321 tagattcata caagagccgt acgttgctct cgtaccagaa gacataccag cgggatcaat
     1381 tatcttcaaa gtccatgctg tcgaccgaga tactggatcg ggtgggtctg tcacctattt
     1441 cctacagaat ctccatagtc cttttgcggt agatcgtcat agtggcgtcc tcagactcca
     1501 agcgggagct accttggact acgaaaggtc ccgcacccac tatataaccg ttgtcgcaaa
     1561 agacgggggt gggaggctgc acggcgcaga cgtggtattt tcagccacaa ccacggtcac
     1621 tgtgaacgtg gaggacgtac aggatatggc cccggtattt gtgggcacac cgtattacgg
     1681 gtatgtttat gaagacacct taccaggttc cgaagtactc aaggtggttg ctatggatgg
     1741 agatagaggc aaaccgaatc gcattcttta ctcactcgtg aatggcaacg acggggcatt
     1801 cgagattaac gagacgtcgg gggcgatatc gatcacccaa agtccagcac agcttcaaag
     1861 agaagtctac gaattacatg ttcaagtaac tgagatgagc cctgcgggct cgcctgctgc
     1921 gcaagctacc gtcccggtga caattagaat cgtggacctg aacaaccatc ccccgacgtt
     1981 ttacggagaa agtggaccac agaatagatt tgaattgagt atgaatgaac accccccgca
     2041 aggagagatt ctgcgaggcc tcaaaattac ggtaaatgat tcagaccagg gagccaatgc
     2101 gaagtttaat cttcaacttg tcgggccgcg aggtattttt cgtgtagtac cacagactgt
     2161 gctgaacgag gcccaggtca ccattatagt cgaaaatagc gcggccatcg atttcgaaaa
     2221 atcaaaggtg ctcacattca agttattagc ggtagaagta aatacgccgg aaaaattctc
     2281 ttcgacggct gacgtcgtga ttcaactttt ggacacaaac gataacgtac ctaaatttga
     2341 ctcgctgtat tacgtggcga ggatacctga aaacgctcca ggcggatctt cagtggtagc
     2401 cgttacagcg gttgacccag acacgggacc ctggggggag gtgaagtaca gcacctatgg
     2461 cacgggcgcg gatctcttct tgattcaccc gagtaccggc ttaatctata cgcaaccttg
     2521 ggcttctcta gacgccgaag ccactgcacg atacaacttc tacgtaaaag cggaagacat
     2581 ggaagggaag tactcagtgg ctgaggtgtt cattacccta ctcgatgtta atgaccaccc
     2641 tccccagttc ggaaagagcg tgcaaaaaaa aacaatggtt ctaggaactc ctgttaagat
     2701 agaagcaatt gatgaagatg ctgaagaacc caacaatcta gttgattact cgatcacgca
     2761 cgcggagcca gcaaatgtat ttgatatcaa cagtcacaca ggggaaatat ggctgaaaaa
     2821 tagtataagg tccctcgatg cactacataa tataacaccc ggacgtgact gtctatggtc
     2881 ccttgaggtc caggccaaag atcgcggctc tccatccttc tctacaactg cactgctgaa
     2941 gatagacatc actgatgcag agactctttc gcgctcacct atggctgcct ttttaataca
     3001 gactaaggat aaccccatga aagcggtcgg cgttttggcc ggtactatgg caaccgttgt
     3061 ggcgatcaca gttttgatca gtacagccac tttctggcgg aacaagaaaa gcaacaaggt
     3121 attgccgatg cggcgtgtct tacgcaagag gccgagtcct gcaccccgaa ctattcgtat
     3181 cgagtggctc aagtcaaaat ctacgaaggc agccacaaag ttcatgttaa aggagaagcc
     3241 cccaaacgaa aattgcaaca acaactctcc tgagtcatcc ctactaccaa gagctccagc
     3301 cttgcccccg cccccctcgg ttgcacccag caccggcgct gcacagtgga ctgtacctac
     3361 ggtcagcgga agtctaacgc cgcagcctac ccagcctccc cctaagccta agacgatggg
     3421 gtctccagtt caaagcacac tgatctctga gctaaaacaa aaattcgaga aaaagtctgt
     3481 gcacaataag gcctattttg gtggtggtgg aagtgaaaat ctttattttc aaggtggagg
     3541 ggggggctcc gcagagcaga agcttataag cgaggaggac cttggcggcg gtgggtccga
     3601 ctacaaagac catgatggcg attataaaga tcatgacatt gattataaag acgatgatga
     3661 caaaggaggt ggtgggtccc atcatcacca tcaccatcat catcaccact gagcggccgc
     3721 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     3781 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3841 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3901 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3961 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     4021 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4081 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4141 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4201 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4261 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4321 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4381 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4441 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4501 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4561 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4621 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4681 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4741 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4801 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4861 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4921 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4981 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5041 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5101 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5161 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5221 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5281 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5341 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5401 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5461 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5521 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5581 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5641 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5701 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5761 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5821 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5881 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5941 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6001 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6061 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6121 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6181 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6241 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6301 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6841 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6961 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7021 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7081 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7141 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7201 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7261 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7321 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7381 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7441 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7501 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7561 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7621 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7681 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7741 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7801 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7861 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7921 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7981 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8041 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8101 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8161 gacgtc
//
