LOCUS       sequence-248281-        6701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of hArl8b with C-term GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6701)
  AUTHORS   Kerr SC, Kahn RA
  TITLE     Tool box: Plasmids for the expression or knockdown of human ARF 
            Family GTPases (ARF/ARL/SAR) and their co-expression in bacteria 
            with N-myristoyltransferases.
  JOURNAL   Cell Logist. 2015 Sep 21;5(3):e1090523. doi: 
            10.1080/21592799.2015.1090523. eCollection 2015 Jul-Sep.
  PUBMED    27057421
REFERENCE   2  (bases 1 to 6701)
  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: "Cell 
            Logist."; date: "2015-09-21"; volume: "5(3):e1090523. doi"; pages: "
            10.1080/21592799.2015.1090523. eCollection 2015 Jul-Sep"
COMMENT     Sequence Label: pDEST47-ARL8B-GFP
FEATURES             Location/Qualifiers
     source          1..6701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    680..704
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1280..1304)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1315..1317,1318..1320,1321..2034)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1348..1376)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1971..1991
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2056..2073)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2062..2286
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2332..2760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2419..2438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2629..2650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2769..2789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2774..3103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2954..3089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3016..3035
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3170..3964
                     /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(3224..3243)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3834..3853
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4140..4261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4177..4196)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4231..4250
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4310..4326)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4310..4326)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4323..4345)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4334..4350
                     /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(4358..4364,4365..4382,4383..4388))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4403..4424
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4541..4558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4712..5300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4792..4811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5471..6262,6263..6331))
                     /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     6094..6113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6332..6436)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6511..6530)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttgatcaaa caagtttgta caaaaaagca ggcttagaag gagataaacc
      721 atgctggcgc tcatctcccg cctgctggac tggttccgtt cgctcttctg gaaggaagag
      781 atggagctga cgctcgtggg gctgcagtac tcgggcaaga ccaccttcgt caatgtcatc
      841 gcgtcaggtc aattcagtga agatatgata cccacagtgg gcttcaacat gaggaaggta
      901 actaaaggta acgtcacaat aaagatctgg gacataggag gacaaccccg atttcgaagc
      961 atgtgggagc ggtattgcag aggagtcaat gctattgttt acatgataga tgctgcagat
     1021 cgtgaaaaga tagaagcttc ccgaaatgag ctacataatc ttctagataa accacagtta
     1081 caaggaattc cagtgctagt gcttggaaac aagagagatc ttcctaatgc cttggatgag
     1141 aaacagctaa ttgaaaaaat gaatctgtct gctattcagg atagagaaat ttgctgctat
     1201 tcaatttctt gcaaagaaaa ggataatata gatatcacac ttcagtggct tattcagcat
     1261 tcaaaatcta gaagaagcga cccagctttc ttgtacaaag tggttcgatc tagaatggct
     1321 agcaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
     1381 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgctac atacggaaag
     1441 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc
     1501 actactttct cttatggtgt tcaatgcttt tcccgttatc cggatcatat gaaacggcat
     1561 gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat atctttcaaa
     1621 gatgacggga actacaagac gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     1681 cgtatcgagt taaaaggtat tgattttaaa gaagatggaa acattctcgg acacaaactc
     1741 gagtacaact ataactcaca caatgtatac atcacggcag acaaacaaaa gaatggaatc
     1801 aaagctaact tcaaaattcg ccacaacatt gaagatggat ccgttcaact agcagaccat
     1861 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
     1921 tcgacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttctt
     1981 gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa ataatgaatt
     2041 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2101 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2161 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2221 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2281 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2341 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2401 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2461 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2521 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2581 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2641 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2701 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2761 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2821 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2881 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2941 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3001 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3061 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3121 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3181 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3241 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3301 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3361 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3421 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3481 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3541 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3601 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3661 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     3721 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3781 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3841 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3901 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3961 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4021 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4081 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4141 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4201 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4261 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4321 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4381 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4441 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4501 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4561 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4621 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4681 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4741 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4801 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4861 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4921 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4981 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5041 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5101 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5161 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5221 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5281 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5341 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5401 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5461 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5521 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5581 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5641 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5701 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5761 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5821 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5881 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5941 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6001 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6061 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6121 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6181 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6241 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6301 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6361 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6421 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     6481 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6541 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     6601 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     6661 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
