LOCUS       sequence-403824-        5168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for bacterial expression of human protein ARF6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5168)
  TITLE     Kahn Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5168)
  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-ARF6-His6(CO)
FEATURES             Location/Qualifiers
     source          1..5168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(404..451)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     447..465
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(519..536)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1087..1109
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1140..1159)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1141..1159)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1243..1262)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             2720..2911
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2905..2927)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3013..3155
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3068..3087
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3170..3187)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3341..3929)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3421..3440)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4100..4891,4892..4960))
                     /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     4723..4742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4961..5065)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5133..5151
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
      241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
      301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
      361 cacacccgtc ctgtggatat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga
      421 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact
      481 cagcttcctt tcgggctttg ttagcagccg gatccttaat gatggtggtg atgatggctg
      541 cccgaaccgc tgccgctttt atagttgctg gtcagccagg tcaggccttc atacaggcca
      601 tcgccgctgg tcgcgcagct cggctgcacg taccaattac gatcgcgaat gcgggtcagg
      661 cccagttttt cctgaatttc gtgcggtttc atggcatccg gcagatcctg tttgttggca
      721 aaaatcagaa tgatcgcatc acgcatttca cgatcgttaa tgatgcggtg cagttcctgg
      781 cgcgcttcat cgatgcgatc acgatccgcg caatccacca caaaaatcag gccctgggtg
      841 ccggtataat agtgacgcca cagcgggcga attttatcct ggccacctac atcccacacg
      901 ttaaatttca cgtttttata cgtcacggtt tccacgttga agcccacggt cggaatcgtg
      961 gtcacgctct ggcccagttt cagtttatac agaatggtgg ttttgcccgc cgcgtccagg
     1021 cccagcatca gaatgcgcat ttctttattg ccaaaaattt tgctcagcac tttgcccata
     1081 tgtatatctc cttcttaaag ttaaacaaaa ttatttctag agggaaaccg ttgtggtctc
     1141 cctatagtga gtcgtattaa tttcgcggga tcgagatctc gatcctctac gccggacgca
     1201 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     1261 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     1321 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     1381 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
     1441 cgcataaggg agagcgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc
     1501 ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac
     1561 tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga
     1621 gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag
     1681 ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca
     1741 tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct
     1801 tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc
     1861 tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta
     1921 ccagcctaac ttcgatcact ggaccgctga tcgtcacggc gatttatgcc gcctcggcga
     1981 gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc tgcctccccg
     2041 cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct
     2101 cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa
     2161 tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac
     2221 gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc
     2281 gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat
     2341 acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg
     2401 aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac
     2461 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
     2521 tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca
     2581 taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac
     2641 ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc
     2701 cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
     2761 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
     2821 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
     2881 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2941 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3001 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     3061 aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg tgtgaaatac
     3121 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3181 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3241 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3301 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3361 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3421 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3481 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3541 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3601 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3661 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3721 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3781 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3841 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3901 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3961 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4021 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4081 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4141 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4201 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4261 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4321 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4381 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
     4441 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4501 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4561 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4621 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4681 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     4741 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4801 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4861 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4921 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4981 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5041 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5101 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5161 ttcaagaa
//
