LOCUS       sequence-402993-        5195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for bacterial expression of human protein ARF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5195)
  TITLE     Kahn Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5195)
  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-ARF1-His6
FEATURES             Location/Qualifiers
     source          1..5195
                     /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             723..740
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(794..812)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      808..855
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1221..1226,1227..1243,1244..1249))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1276..1294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1362..1466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1467..1535,1536..2327)
                     /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(1685..1704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2498..3086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2987..3006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3240..3257
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3272..3414
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3340..3359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3500..3522
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3516..3707)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5165..5184
                     /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 atatggggct cctgaccatt ctgaagaaga
      181 tgaagcagaa agagcgggag ctgcgactgc tcatgcttgg cctggacaat gctggaaaga
      241 caaccatcct gaagaagttc aatggggagg acatcgacac catctcccca acgctgggct
      301 tcaacatcaa gaccctggag caccgaggat tcaagctgaa catctgggat gtgggtggcc
      361 agaagtccct gcggtcctac tggcggaact actttgagag caccgatggc ctcatctggg
      421 tagtggacag cgcagaccgc cagcgcatgc aggactgcca gcgggagctc cagagcctgc
      481 tggtggagga gcgcctggcc ggagcaaccc tcctcatctt tgctaataag caggacctgc
      541 ctggagcact gtcctctaac gccatccgcg aggtcctgga gctggactcc atccgcagcc
      601 accactggtg catccagggc tgcagcgccg tcaccgggga gaacctgctg ccgggcatcg
      661 actggctcct ggatgacatt tccagccgca ttttcacagc tgacggcagc ggcagcggca
      721 gccatcacca tcaccatcac taaggatccg gctgctaaca aagcccgaaa ggaagctgag
      781 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
      841 ttgaggggtt ttttgctgaa aggaggaact atatccggat atccacagga cgggtgtggt
      901 cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg
      961 gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat agaaattgca tcaacgcata
     1021 tagcgctagc agcacgccat agtgactggc gatgctgtcg gaatggacga tatcccgcaa
     1081 gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc
     1141 gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt
     1201 taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt
     1261 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1321 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1381 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1441 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1501 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1561 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1621 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1681 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     1741 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1801 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1861 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1921 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1981 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2041 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2101 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2161 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2221 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2281 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2341 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2401 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2461 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2521 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2581 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2641 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2701 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2761 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2821 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2881 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2941 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3001 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3061 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3121 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3181 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3241 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3301 tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc
     3361 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3421 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3481 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3541 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     3601 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     3661 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     3721 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     3781 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     3841 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     3901 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     3961 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4021 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     4081 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4141 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg
     4201 cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt
     4261 tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg
     4321 ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac
     4381 gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc
     4441 atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccagtg atcgaagtta
     4501 ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc
     4561 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa
     4621 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg
     4681 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     4741 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     4801 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     4861 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     4921 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga cgctctccct
     4981 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     5041 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     5101 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     5161 catcggtgat gtcggcgata taggcgccag caacc
//
