LOCUS       sequence-267708-        7525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human elF4A1 residues 1-406, N-term His tag, thrombin cleavage, in 
            pET21 vector backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7525)
  TITLE     Wagner Lab plasmids
REFERENCE   2  (bases 1 to 7525)
  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: KE-005
FEATURES             Location/Qualifiers
     source          1..7525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     126..145
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        126..144
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    145..169
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             184..206
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             225..242
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             252..269
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2152..2170)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2166..2213
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2388..2393,2394..2410,2411..2416))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2443..2461)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2529..2633
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2634..2702,2703..3494)
                     /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(2852..2871)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3665..4253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4154..4173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4407..4424
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4439..4578
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4504..4523)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4664..4686
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4680..4871)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     7226..7245
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7265..7342)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
       61 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
      121 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
      181 aattttgttt aactttaaga aggagatata ccatgggcag cagccatcat catcatcatc
      241 acagcagcgg cctggtgccg cgcggcagcc atatgctcga ggatccgttt ctaaggatca
      301 tgtctgcgag ccaggattcc cgatccagag acaatggccc cgatgggatg gagcccgaag
      361 gcgtcatcga gagtaactgg aatgagattg ttgacagctt tgatgacatg aacctctcgg
      421 agtcccttct ccgtggcatc tacgcctatg gttttgagaa gccctctgcc atccagcagc
      481 gagccattct accttgtatc aagggttatg atgtgattgc tcaagcccaa tctgggactg
      541 ggaaaacggc cacatttgcc atatcgattc tgcagcagat tgaattagat ctaaaagcca
      601 cccaggcctt ggtcctagca cccactcgag aattggctca gcagatacag aaggtggtca
      661 tggcactagg agactacatg ggcgcctcct gtcacgcctg tatcgggggc accaacgtgc
      721 gtgctgaggt gcagaaactg cagatggaag ctccccacat catcgtgggt acccctggcc
      781 gtgtgtttga tatgcttaac cggagatacc tgtcccccaa atacatcaag atgtttgtac
      841 tggatgaagc tgacgaaatg ttaagccgtg gattcaagga ccagatctat gacatattcc
      901 aaaagctcaa cagcaacacc caggtagttt tgctgtcagc cacaatgcct tctgatgtgc
      961 ttgaggtgac caagaagttc atgagggacc ccattcggat tcttgtcaag aaggaagagt
     1021 tgaccctgga gggtatccgc cagttctaca tcaacgtgga acgagaggag tggaagctgg
     1081 acacactatg tgacttgtat gaaaccctga ccatcaccca ggcagtcatc ttcatcaaca
     1141 cccggaggaa ggtggactgg ctcaccgaga agatgcatgc tcgagatttc actgtatccg
     1201 ccatgcatgg agatatggac caaaaggaac gagacgtgat tatgagggag tttcgttctg
     1261 gctctagcag agttttgatt accactgacc tgctggccag aggcattgat gtgcagcagg
     1321 tttctttagt catcaactat gaccttccca ccaacaggga aaactatatc cacagaatcg
     1381 gtcgaggtgg acggtttggc cgtaaaggtg tggctattaa catggtgaca gaagaagaca
     1441 agaggactct tcgagacatt gagaccttct acaacacctc cattgaggaa atgcccctca
     1501 atgttgctga cctcatctga ggggctgtcc tgccacccag ccccagccag ggctcaatct
     1561 ctgggggctg aggagcagca ggagggggga gggaagggag ccaagggatg gacatcttgt
     1621 catttttttt ctttgaataa atgtcacttt ttgaggcaaa agaaggaaaa aaaaaaaaaa
     1681 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1741 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1801 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1861 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1921 aaaraaaaaa araaaaaaaa aaaaaaaaaa aananaaaaa aaaaaaaaaa aaaaaaaaaa
     1981 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2041 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaag taccgcggcc
     2101 gcgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     2161 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     2221 gaggaactat atccggatat cccgcaagag gcccggcagt accggcataa ccaagcctat
     2281 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
     2341 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc
     2401 gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat
     2461 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2521 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2581 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2641 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2701 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2761 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2821 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     2881 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2941 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3001 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3061 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3121 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     3181 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3241 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3301 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3361 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3421 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3481 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3541 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3601 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3661 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3721 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3781 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3841 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3901 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3961 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4021 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4081 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4141 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4201 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4261 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4321 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4381 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4441 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     4501 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     4561 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4621 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4681 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     4741 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     4801 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     4861 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     4921 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     4981 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     5041 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     5101 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     5161 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     5221 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     5281 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     5341 caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg
     5401 gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct
     5461 ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg
     5521 tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc
     5581 ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga
     5641 tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc
     5701 cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc
     5761 cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca
     5821 gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga
     5881 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     5941 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     6001 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     6061 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     6121 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     6181 gctcactgcc cgctttccag tcgggaaact gtcgtgccag ctgcattaat gaatcggcca
     6241 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     6301 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     6361 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     6421 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     6481 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     6541 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     6601 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     6661 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     6721 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     6781 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     6841 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     6901 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     6961 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     7021 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     7081 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     7141 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     7201 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     7261 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     7321 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     7381 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     7441 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     7501 aaacaagcgc tcatgagccc gaagt
//
