LOCUS       sequence-403014-        5189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for bacterial expression of human protein ARL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5189)
  TITLE     Kahn Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5189)
  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"
COMMENT     Sequence Label: pET3-ARL3-His6
FEATURES             Location/Qualifiers
     source          1..5189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..92
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        73..91
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             123..145
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             717..734
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(788..806)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      802..849
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1215..1220,1221..1237,1238..1243))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1270..1288)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1356..1460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1461..1529,1530..2321)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1679..1698)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2492..3080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2981..3000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3234..3251
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3266..3408
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3334..3353)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3494..3516
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3510..3701)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5159..5178
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
       61 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata
      121 attttgttta actttaagaa ggagatatac atatgggctt gctctcaatt ttgcgcaagt
      181 tgaaaagtgc accagaccag gaggtgagaa tacttctcct gggcttggat aatgctggca
      241 agaccactct tctgaagcag cttgcatctg aagacatcag ccacatcaca cctacacagg
      301 gtttcaacat caaaagtgta caatcacaag gttttaaact gaatgtatgg gacattggtg
      361 gacagaggaa aatcagacca tactggaaga attattttga aaataccgat attcttatat
      421 atgtaatcga cagtgcagac agaaaaagat ttgaagagac gggtcaggaa ctagcggaat
      481 tactggagga agaaaaacta agttgtgtgc cagtgctcat ctttgctaat aagcaggatt
      541 tgctcacagc agcccctgcc tctgaaattg cagaaggact gaacctgcat accatccgcg
      601 accgagtctg gcagatccag tcttgctcag ctctcacagg agagggcgtt caggatggca
      661 tgaactgggt ctgcaaaaat gtcaatgcaa agaagaaagg cagcggcagc ggcagccatc
      721 accatcacca tcactaagga tccggctgct aacaaagccc gaaaggaagc tgagttggct
      781 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
      841 ggttttttgc tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat
      901 gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa
      961 gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc
     1021 tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc
     1081 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
     1141 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
     1201 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
     1261 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1321 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1381 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1441 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1501 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1561 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1621 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1681 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     1741 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1801 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1861 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1921 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1981 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     2041 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2101 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2161 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2221 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2281 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2341 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2401 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2461 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2521 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2581 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2641 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2701 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2761 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2821 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2881 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2941 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3001 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3061 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3121 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3181 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3241 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     3301 gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3361 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     3421 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3481 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3541 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     3601 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     3661 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     3721 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     3781 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     3841 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     3901 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     3961 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     4021 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     4081 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     4141 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc
     4201 gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca
     4261 ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg
     4321 aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg
     4381 cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc
     4441 tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc agtgatcgaa gttaggctgg
     4501 taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca
     4561 gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga
     4621 aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca
     4681 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     4741 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     4801 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     4861 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     4921 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgacgctct cccttatgcg
     4981 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     5041 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     5101 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     5161 tgatgtcggc gatataggcg ccagcaacc
//
