LOCUS       sequence-269573-        5248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heat 3 domain of human elF4G in pET21 backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5248)
  TITLE     Wagner Lab plasmids
REFERENCE   2  (bases 1 to 5248)
  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: RG-001
FEATURES             Location/Qualifiers
     source          1..5248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        37..55
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    83..107
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             119..139
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             152..172
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             173..193
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(733..757)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      824..871
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1237..1242,1243..1259,1260..1265))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1292..1310)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1378..1482
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1483..1551,1552..2343)
                     /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(1701..1720)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2514..3102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3003..3022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3256..3273
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3288..3427
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3353..3372)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3513..3535
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3529..3720)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5182..5201
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact atagggagac
       61 cacaacggtt tccctctaga tcacaagttt gtacaaaaaa gcaggctccg cggccgcctt
      121 gtttaacttt aagaaggagc ccttcacaat gcaccatcac catcaccatc acgaaaacct
      181 gtattttcag ggtgaggaat cggaagcccc tggccagagg gcactcccct ccgaggagct
      241 gaacaggcag ctggagaagc tgctgaagga gggcagcagt aaccagcggg tgttcgactg
      301 gatagaggcc aacctgagtg agcagcagat agtatccaac acgttagttc gagccctcat
      361 gacggctgtc tgctattctg caattatttt tgagactccc ctccgagtgg acgttgcagt
      421 gctgaaagcg cgagcgaagc tgctgcagaa atacctgtgt gacgagcaga aggagctaca
      481 ggcgctctac gccctccagg cccttgtagt gaccttagaa cagcctccca acctgctgcg
      541 gatgttcttt gacgcactgt atgacgagga cgtggtgaag gaggatgcct tctacagttg
      601 ggagagtagc aaggaccccg ctgagcagca gggcaagggt gtggccctta aatctgtcac
      661 agccttcttc aagtggctcc gtgaagcaga ggaggagtct gaccacaact gaattcaagg
      721 gtgggcgcgc cgacccagct ttcttgtaca aagtggtgat gatccggctg ctaacaaagc
      781 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
      841 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc
      901 acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga
      961 gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa
     1021 attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat
     1081 ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     1141 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     1201 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     1261 gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     1321 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     1381 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1441 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1501 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1561 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1621 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1681 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     1741 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1801 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1861 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1921 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1981 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     2041 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2101 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2161 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2221 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2281 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2341 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2401 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2461 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2521 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2581 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2641 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2701 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2761 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2821 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2881 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2941 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3001 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3061 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3121 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3181 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3241 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3301 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     3361 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     3421 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3481 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3541 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     3601 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     3661 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     3721 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     3781 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     3841 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     3901 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     3961 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     4021 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     4081 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     4141 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac
     4201 gctgcccgag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg
     4261 ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg
     4321 agtggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc
     4381 catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat
     4441 gccaacccgt tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca
     4501 atgatcgaag ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg
     4561 tcatctacct gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg
     4621 agaagaatca taatggggaa ggccatccag cctcgcgtcg cgaacgccag caagacgtag
     4681 cccagcgcgt cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     4741 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     4801 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     4861 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     4921 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     4981 cgatcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     5041 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     5101 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     5161 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     5221 ggcgccggtg atgccggcca cgatgcgt
//
