LOCUS       sequence-403021-        5201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for bacterial expression of human protein ARL8A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5201)
  TITLE     Kahn Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5201)
  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-ARL8A-His6
FEATURES             Location/Qualifiers
     source          1..5201
                     /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             729..746
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(800..818)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      814..861
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1227..1232,1233..1249,1250..1255))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1282..1300)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1368..1472
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1473..1541,1542..2333)
                     /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(1691..1710)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2504..3092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2993..3012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3246..3263
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3278..3420
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3346..3365)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3506..3528
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3522..3713)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5171..5190
                     /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 atatgatcgc tttgttcaac aagctgctgg
      181 actggttcaa ggccctattc tggaaggagg agatggagct cacgctggtc gggcttcagt
      241 actcgggcaa gaccaccttc gtcaacgtga tcgcgtcagg acagttcaac gaggacatga
      301 tccccaccgt gggtttcaac atgcgcaaaa tcaccaaagg gaatgtgact atcaagctct
      361 gggacattgg gggacagccg cgtttccgca gcatgtggga gcgctactgc cgaggagtga
      421 gcgccatcgt gtacatggtg gatgctgctg accaggagaa gattgaggcc tctaagaacg
      481 agctccacaa cctactggac aaacctcagc tgcagggcat cccggtctta gtcctgggta
      541 acaagcgaga ccttccggga gcattggatg agaaggagct gattgagaaa atgaatctgt
      601 ctgccatcca ggaccgagag atctgctgct actccatctc ttgcaaagaa aaggacaaca
      661 ttgacatcac cctacagtgg cttattcaac actcgaagtc acggagaagc ggcagcggca
      721 gcggcagcca tcaccatcac catcactaag gatccggctg ctaacaaagc ccgaaaggaa
      781 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      841 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc acaggacggg
      901 tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga gcaggactgg
      961 gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa attgcatcaa
     1021 cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat ggacgatatc
     1081 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     1141 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
     1201 gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg
     1261 agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     1321 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1381 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1441 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1501 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1561 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1621 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1681 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     1741 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1801 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1861 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1921 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1981 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     2041 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2101 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2161 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2221 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2281 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2341 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2401 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2461 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2521 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2581 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2641 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2701 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2761 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2821 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2881 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2941 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3001 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3061 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3121 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3181 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3241 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3301 cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga
     3361 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     3421 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3481 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3541 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     3601 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     3661 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     3721 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     3781 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     3841 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     3901 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     3961 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     4021 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     4081 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     4141 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
     4201 gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg
     4261 ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg
     4321 aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac
     4381 cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc
     4441 cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccagtgatcg
     4501 aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta
     4561 cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa
     4621 tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg
     4681 cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
     4741 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
     4801 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
     4861 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     4921 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgacgct
     4981 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     5041 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     5101 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     5161 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac c
//
