LOCUS       sequence-414034-        5934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human LARP1 LaM domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5934)
  AUTHORS   Kozlov G, Mattijssen S, Jiang J, Nyandwi S, Sprules T, Iben JR, Coon
            SL, Gaidamakov S, Noronha AM, Wilds CJ, Maraia RJ, Gehring K
  TITLE     Structural basis of 3'-end poly(A) RNA recognition by LARP1.
  JOURNAL   Nucleic Acids Res. 2022 Sep 9;50(16):9534-9547. doi: 
            10.1093/nar/gkac696.
  PUBMED    35979957
REFERENCE   2  (bases 1 to 5934)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2022-09-9- 9"; volume: "50"; issue: 
            "16"; pages: "9534-9547"
COMMENT     Sequence Label: LARP1_323-410
FEATURES             Location/Qualifiers
     source          1..5934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(839..856)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             875..897
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    912..936
                     /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)."
     primer_bind     complement(936..955)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(937..955)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1039..1058)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1264..1341
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1342..2424
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1361..1380)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3233..3424
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3418..3440)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3526..3665
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3581..3600
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3680..3697)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3851..4439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3931..3950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4550..4569)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4561..5376
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4561..5376
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4634..4653)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5468..5923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5605..5626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5817..5836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gacaagcttt tatgatcctt
      181 aatccatctg ctccatctcc tcgtcaaaca gaaaatccag ttcctcttgc tcatcctgat
      241 ccttggacat cagctgctgg gaaggcagct gctaaggcag ggaggtcagg tgggaaaatc
      301 tggactcctc tgacttcttg ggccgtgctg gggatggccg aggcctcttc ttcacttcaa
      361 tccagttctc agaatccagg tcaggcaggc tggcagacag gcccttgggt agtgtcttta
      421 ggttgctgac ctcctctgtt ttggttggca ctggggtgac tgcacgagga gagccaggtg
      481 ccgactctgt ctccttttgg tagtgctgac ggggaacaaa ttcagggcag ttgagaagct
      541 gggagaaatc agtctgtgaa taatccactt atgggggaag aggccacttt tctggttcct
      601 ccctcctacg aactttctca tcaacgatct ccaccacctt gctgtccttt agggccgcaa
      661 agatgagtga aatgtcagtg gtaagggcct gcactcggtg gaaggaagca ataagggtga
      721 tgggtaggaa accatcagca tccattttcc ttcgcaggaa gaagtctcgc tctaaattgt
      781 ccacgctgaa gtagtattca atctggcgct tgatgtagtc tttgagcagt tcctggctgt
      841 gatgatgatg atgatggctg ctgcccatgg tatatctcct tcttaaagtt aaacaaaatt
      901 atttctagag gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc
      961 gcgggatcga gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     1021 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     1081 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     1141 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
     1201 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc
     1261 ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     1321 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     1381 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     1441 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     1501 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     1561 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     1621 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     1681 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     1741 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     1801 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     1861 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     1921 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     1981 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     2041 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     2101 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     2161 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     2221 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     2281 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     2341 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2401 tcccgactgg aaagcgggca gtgagcacaa cgcaattaat gtaagttagc tcactcatta
     2461 ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     2521 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     2581 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     2641 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     2701 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     2761 ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     2821 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     2881 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     2941 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     3001 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     3061 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     3121 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     3181 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     3241 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     3301 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     3361 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     3421 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     3481 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     3541 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     3601 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     3661 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3721 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3781 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3841 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3901 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3961 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4021 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4081 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4141 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4201 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4261 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4321 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4381 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4441 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4501 ggattttggt catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     4561 atgagccata ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat
     4621 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     4681 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     4741 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     4801 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     4861 atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     4921 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     4981 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     5041 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     5101 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     5161 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     5221 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     5281 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     5341 ttgcagtttc atttgatgct cgatgagttt ttctaagaat taattcatga gcggatacat
     5401 atttgaatgt atttagaaaa ataaacaaat aggggttccg tgcacatttc cccgaaaagt
     5461 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5521 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5581 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5641 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5701 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5761 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5821 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5881 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gcca
//
