LOCUS       sequence-494571-        7307 bp    DNA     circular SYN 04-DEC-2025
DEFINITION  Mammalian expression of truncated Arl13b containing the Proline-Rich
            domain at the C-terminus (cytosol localization)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7307)
  AUTHORS   Liu X, Yam PT, Schlienger S, Cai E, Zhang J, Chen WJ, Torres 
            Gutierrez O, Jimenez Amilburu V, Ramamurthy V, Ting AY, Branon TC, 
            Cayouette M, Gen R, Marks T, Kong JH, Charron F, Ge X
  TITLE     Numb positively regulates Hedgehog signaling at the ciliary pocket.
  JOURNAL   Nat Commun. 2024 Apr 25;15(1):3365. doi: 10.1038/s41467-024-47244-1.
  PUBMED    38664376
REFERENCE   2  (bases 1 to 7307)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2024-04-25"; volume: "15(1):3365. doi"; pages: " 
            10.1038/s41467-024-47244-1"
COMMENT     Sequence Label: pcDNA3-Arl13b-PR-EGFP-TurboID
FEATURES             Location/Qualifiers
     source          1..7307
                     /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      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1111..1113,1114..1116,1117..1827)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1111..1113,1114..1116,1117..1827)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1417..1436)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1764..1785
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1834..2790
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             2797..2823
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     promoter        complement(2860..2878)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2861..2878)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2898..2915)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2904..3128
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3174..3602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3261..3280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3471..3492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3611..3631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3616..3945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3796..3931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3858..3877
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4012..4806
                     /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(4066..4085)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4676..4695
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4980..5101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5017..5036)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5071..5090
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5150..5166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5150..5166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5163..5185)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5174..5190
                     /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(5198..5204,5205..5222,5223..5228))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5243..5264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5381..5398)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5552..6140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5632..5651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6311..7102,7103..7171))
                     /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"
     promoter        complement(7172..7276)
                     /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 gcttgccacc
      901 atggtcaata cagacgagtc tactccaaag agtcccacgc ctccccaacc tccccctcct
      961 gttggctggg gaacccccaa agtcactaga cttccaaaac ttgagcctct tggtgaaaca
     1021 cgtcacaatg atttctatgg gaagccgctg cctcccctgg ctgtgcgaca gagacctaac
     1081 ggtgatgctc aggacacgat ctcaggatcc atggtgagca agggcgagga gctgttcacc
     1141 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1201 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1261 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1321 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1381 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1441 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1501 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1561 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     1621 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1681 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     1741 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1801 actctcggca tggacgagct gtacaaggct agcaaagaca atactgtgcc tctgaagctg
     1861 atcgctctcc tggctaatgg cgagttccat agtggcgaac agctgggaga aaccctgggc
     1921 atgtccaggg ccgctatcaa caagcacatt cagactctgc gcgactgggg cgtggacgtg
     1981 ttcaccgtgc ccggaaaggg ctactctctg cccgagccta tcccgctgct gaacgctaaa
     2041 cagattctgg gacagctgga cggcgggagc gtggcagtcc tgcctgtggt cgactccacc
     2101 aatcagtacc tgctggatcg aatcggcgag ctgaagagtg gggatgcttg cattgcagaa
     2161 tatcagcagg cagggagagg aagcagaggg aggaaatggt tctctccttt tggagctaac
     2221 ctgtacctga gtatgttttg gcgcctgaag cggggaccag cagcaatcgg cctgggcccg
     2281 gtcatcggaa ttgtcatggc agaagcgctg cgaaagctgg gagcagacaa ggtgcgagtc
     2341 aaatggccca atgacctgta tctgcaggat agaaagctgg caggcatcct ggtggagctg
     2401 gccggaataa caggcgatgc tgcacagatc gtcattggcg ccgggattaa cgtggctatg
     2461 aggcgcgtgg aggaaagcgt ggtcaatcag ggctggatca cactgcagga agcagggatt
     2521 aacctggaca ggaatactct ggccgctacg ctgatccgag agctgcgggc agccctggaa
     2581 ctgttcgagc aggaaggcct ggctccatat ctgccacggt gggagaagct ggataacttc
     2641 atcaatagac ccgtgaagct gatcattggg gacaaagaga ttttcgggat tagccggggg
     2701 attgataaac agggagccct gctgctggaa caggacggag ttatcaaacc ctggatgggc
     2761 ggagaaatca gtctgcggtc tgccgaaaag ctgcagctgc ctcccctgga gcgcctgacc
     2821 ctggactaat agctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     2881 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2941 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3001 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3061 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3121 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3181 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3241 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3301 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3361 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3421 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3481 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3541 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3601 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3901 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3961 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4021 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4081 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4141 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4201 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4261 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4321 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4381 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4441 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4501 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4561 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4621 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4681 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4741 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4801 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4861 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4921 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4981 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5041 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5101 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5161 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5221 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5281 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5341 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5401 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5461 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5521 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5581 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5641 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5701 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5761 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac aaaccccccg
     5821 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5881 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5941 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6001 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6061 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6121 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6181 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6241 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6301 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6361 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6421 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6481 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6541 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6601 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6661 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6721 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6781 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6841 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6901 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6961 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7021 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7081 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7141 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7201 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7261 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
