LOCUS       sequence-420100-        8673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian overexpression vector for C-terminally mNeonGreen tagged 
            human Pchd21 (WT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8673)
  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 8673)
  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-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..8673
                     /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"
     regulatory      3764..3773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4264..4281)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4270..4494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4540..4968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4627..4646)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4837..4858
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4977..4997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4982..5311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5162..5297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5224..5243
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5378..6172
                     /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(5432..5451)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6042..6061
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6346..6467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6383..6402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6437..6456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6516..6532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6516..6532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6529..6551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6540..6556
                     /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(6564..6570,6571..6588,6589..6594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6609..6630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6747..6764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6918..7506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6998..7017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7677..8468,8469..8537))
                     /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     8300..8319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8538..8642)
                     /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 gtggtagtgg agttagcaag ggtgaagaag acaatatggc
     3541 tagcttgccc gccacacacg agctgcacat atttgggtct ataaatggag tggattttga
     3601 tatggtcggc caaggaaccg ggaatcccaa tgacgggtat gaggagctta acctgaaaag
     3661 cacaaaaggc gacttgcagt tttcaccctg gattctggtc ccgcatatcg gatacggctt
     3721 ccatcagtat ctgccatacc ctgatggaat gagccctttt caggccgcca tggtggacgg
     3781 atcaggctac caggtgcatc ggacgatgca atttgaggac ggtgctagcc tgactgttaa
     3841 ttatcgatac acctatgagg ggtctcacat taagggagag gcccaggtca aggggacagg
     3901 atttccagcc gatggtccag tcatgacgaa cagtctgaca gctgccgact ggtgtcgctc
     3961 taaaaaaaca taccccaatg acaaaaccat aatctctacc ttcaagtggt cctacacgac
     4021 aggtaacgga aagagataca ggtccactgc ccggacaacc tataccttcg ccaagcctat
     4081 ggcggctaat tacttgaaga atcagcccat gtatgtgttt agaaagacag agctgaaaca
     4141 ttccaagact gagctgaatt tcaaggagtg gcagaaagct tttacagatg tcatgggtat
     4201 ggatgagctg tacaagtgag cggccgctcg agtctagagg gcccgtttaa acccgctgat
     4261 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4321 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4381 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4441 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4501 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4561 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4621 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4681 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4741 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4801 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4861 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4921 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4981 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5041 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5101 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5161 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5221 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     5281 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     5341 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5401 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5461 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5521 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5581 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5641 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5701 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5761 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5821 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5881 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5941 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6001 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6061 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6121 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6181 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6241 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6301 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6361 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6421 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6481 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6541 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6601 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6661 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6721 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6781 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6841 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6901 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6961 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7021 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7081 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7141 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7201 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7261 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7321 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7381 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7441 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7501 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7561 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7621 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7681 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7741 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7801 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7861 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7921 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7981 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8041 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8101 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8161 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8221 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8281 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8341 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8401 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8461 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8521 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8581 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8641 cgcacatttc cccgaaaagt gccacctgac gtc
//
