LOCUS       sequence-358808-        5158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For purification (from E.coli) of His-tagged TARG1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5158)
  AUTHORS   Zaja R, Aydin G, Lippok BE, Feederle R, Luscher B, Feijs KLH
  TITLE     Comparative analysis of MACROD1, MACROD2 and TARG1 expression, 
            localisation and interactome.
  JOURNAL   Sci Rep. 2020 May 19;10(1):8286. doi: 10.1038/s41598-020-64623-y.
  PUBMED    32427867
REFERENCE   2  (bases 1 to 5158)
  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"; journalName: "Sci Rep."; 
            date: "2020-05-19"; volume: "10(1):8286. doi"; pages: " 
            10.1038/s41598-020-64623-y"
COMMENT     Sequence Label: pDEST17-TARG1
FEATURES             Location/Qualifiers
     source          1..5158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        23..41
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             73..95
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             115..132
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    142..166
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(629..653)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(706..724)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      720..767
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1133..1138,1139..1155,1156..1161))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1188..1206)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1274..1378
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1379..1447,1448..2239)
                     /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(1597..1616)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2410..2998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2899..2918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3152..3169
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3184..3323
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3249..3268)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3409..3431
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3425..3616)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5078..5097
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgagatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc
       61 tctagaaata attttgttta actttaagaa ggagatatac atatgtcgta ctaccatcac
      121 catcaccatc acctcgaatc aacaagtttg tacaaaaaag caggcttcat ggccagcagc
      181 cttaatgaag atccagaagg aagcagaatc acttatgtga aaggagacct ttttgcatgc
      241 ccgaaaacag actctttagc ccactgtatc agtgaggatt gtcgcatggg cgctgggata
      301 gctgtcctct ttaagaagaa atttggaggg gtgcaagaac ttttaaatca acaaaagaaa
      361 tctggagaag tggctgttct gaagagagat gggcgatata tatattactt gattacaaag
      421 aaaagggctt cgcacaagcc aacttatgaa aacttacaga agagtttaga ggcaatgaag
      481 tctcattgtc tgaagaatgg agtcactgac ctctccatgc ccaggattgg atgtggtctt
      541 gatcgtctgc aatgggaaaa tgtatctgcg atgatcgagg aggtatttga ggcaacagac
      601 atcaaaatta ctgtgtacac actctgagac ccagctttct tgtacaaagt ggttgattcg
      661 aggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
      721 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
      781 ctatatccgg atatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct
      841 ccaagtagcg aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga
      901 acgggtgcgc atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg
      961 gcgatgctgt cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag
     1021 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
     1081 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
     1141 ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg
     1201 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     1261 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1321 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1381 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1441 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1501 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1561 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1621 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1681 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1741 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1801 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1861 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1921 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1981 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2041 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2101 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2161 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2221 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2281 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2341 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2401 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2461 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2521 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2581 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2641 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2701 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2761 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2821 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2881 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2941 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3001 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3061 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3121 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3181 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     3241 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     3301 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     3361 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3421 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3481 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3541 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3601 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     3661 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     3721 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     3781 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     3841 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     3901 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     3961 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     4021 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     4081 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
     4141 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
     4201 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
     4261 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
     4321 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
     4381 gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
     4441 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
     4501 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
     4561 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     4621 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     4681 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     4741 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     4801 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4861 tctcaagggc atcggtcgat cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     4921 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     4981 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     5041 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     5101 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggat
//
