LOCUS       sequence-403150-        5183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for bacterial expression of human protein ARF5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5183)
  TITLE     Kahn Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5183)
  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-ARF5-His6(CO)
FEATURES             Location/Qualifiers
     source          1..5183
                     /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             711..728
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(782..800)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      796..843
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1209..1214,1215..1231,1232..1237))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1264..1282)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1350..1454
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1455..1523,1524..2315)
                     /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(1673..1692)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2486..3074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2975..2994
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3228..3245
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3260..3402
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3328..3347)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3488..3510
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3504..3695)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5153..5172
                     /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 atatgggcct caccgtgtcc gcgctctttt
      181 cgcggatctt cgggaagaag cagatgcgga ttctcatggt tggcttggat gcggctggca
      241 agaccacaat cctgtacaaa ctgaagttgg gggagattgt caccaccatc ccaaccatag
      301 gcttcaatgt agaaacagtg gaatataaga acatctgttt cacagtctgg gacgtgggag
      361 gccaggacaa gattcggcct ctgtggcggc actacttcca gaacactcag ggcctcatct
      421 ttgtggtgga cagtaatgac cgggagcggg tccaagaatc tgctgatgaa ctccagaaga
      481 tgctgcagga ggacgagctg cgggatgcag tgctgctggt atttgccaac aagcaggaca
      541 tgcccaacgc catgcccgtg agcgagctga ctgacaagct ggggctacag cacttacgca
      601 gccgcacgtg gtatgtccag gccacctgtg ccacccaagg cacaggtctg tacgatggtc
      661 tggactggct gtcccacgag ctgtcaaagc gcggcagcgg cagcggcagc catcaccatc
      721 accatcacta aggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
      781 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      841 ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc
      901 gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc
      961 ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag
     1021 cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag
     1081 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     1141 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     1201 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
     1261 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1321 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1381 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1441 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1501 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1561 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1621 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1681 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1741 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1801 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1861 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1921 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1981 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2041 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2101 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2161 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2221 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2281 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2341 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2401 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2461 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2521 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2581 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2641 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2701 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2761 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2821 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2881 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2941 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3001 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3061 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3121 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3181 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3241 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3301 cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3361 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     3421 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3481 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     3541 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     3601 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     3661 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     3721 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     3781 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     3841 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     3901 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     3961 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4021 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4081 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     4141 agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg
     4201 ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca
     4261 gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc
     4321 cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga
     4381 ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg
     4441 aggcggcata aatcgccgtg acgatcagcg gtccagtgat cgaagttagg ctggtaagag
     4501 ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
     4561 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
     4621 tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg
     4681 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     4741 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     4801 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     4861 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     4921 agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctta tgcgactcct
     4981 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     5041 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     5101 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     5161 cggcgatata ggcgccagca acc
//
