LOCUS       sequence-403002-        5246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for bacterial expression of human protein ARFRP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5246)
  TITLE     Kahn Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5246)
  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-ARFRP1-His6
FEATURES             Location/Qualifiers
     source          1..5246
                     /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             774..791
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(845..863)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      859..906
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1272..1277,1278..1294,1295..1300))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1327..1345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1413..1517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1518..1586,1587..2378)
                     /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(1736..1755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2549..3137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3038..3057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3291..3308
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3323..3465
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3391..3410)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3551..3573
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3567..3758)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5216..5235
                     /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 atatgtacac gctgctgtcg ggcttgtaca
      181 agtacatgtt tcagaaggac gagtactgca tcctgatcct gggcctggac aatgctggga
      241 agacgacctt cctggagcag tcgaaaaccc gatttaacaa gaactacaag gggatgagtc
      301 tatccaaaat caccaccacc gtgggcctaa acatcggcac tgtggatgtg ggaaaggctc
      361 ggctcatgtt ctgggactta ggagggcagg aagagctgca gtctttgtgg gacaagtatt
      421 atgcggagtg tcacggcgtc atctacgtca ttgactccac cgacgaggag aggctggctg
      481 agtccaagca ggcgtttgag aaggtggtga ccagcgaggc gctgtgcggt gtccccgtct
      541 tggtgctggc caacaagcag gatgtggaga cgtgcctctc aatccctgac atcaagacgg
      601 ccttcagcga ctgcaccagc aagatcggca ggcgagattg cctgacccag gcctgctcgg
      661 ccctcacagg caaaggggtg cgcgagggca tcgagtggat ggtgaagtgt gtcgtgcgga
      721 atgtgcaccg gccgccgcgg cagagggaca tcacgggcag cggcagcggc agccatcacc
      781 atcaccatca ctaaggatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
      841 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
      901 tttttgctga aaggaggaac tatatccgga tatccacagg acgggtgtgg tcgccatgat
      961 cgcgtagtcg atagtggctc caagtagcga agcgagcagg actgggcggc ggccaaagcg
     1021 gtcggacagt gctccgagaa cgggtgcgca tagaaattgc atcaacgcat atagcgctag
     1081 cagcacgcca tagtgactgg cgatgctgtc ggaatggacg atatcccgca agaggcccgg
     1141 cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac
     1201 gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga
     1261 taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat tcttgaagac
     1321 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1381 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1441 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1501 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1561 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1621 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1681 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1741 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     1801 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1861 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1921 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1981 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2041 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2101 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2161 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2221 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2281 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2341 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2401 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     2701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3001 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3061 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3181 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3241 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3301 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3361 tttcacaccg catatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3421 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     3481 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3541 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3601 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     3661 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     3721 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     3781 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     3841 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     3901 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     3961 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     4021 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4081 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4141 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     4201 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     4261 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     4321 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     4381 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     4441 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     4501 ccgaggcggc ataaatcgcc gtgacgatca gcggtccagt gatcgaagtt aggctggtaa
     4561 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     4621 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     4681 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     4741 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4801 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     4861 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     4921 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     4981 aggagctgac tgggttgaag gctctcaagg gcatcggtcg acgctctccc ttatgcgact
     5041 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     5101 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     5161 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     5221 tgtcggcgat ataggcgcca gcaacc
//
