LOCUS       sequence-78371-e        8430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the codon optimized ORF2 of L1RP with his tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8430)
  AUTHORS   Wagstaff BJ, Barnerssoi M, Roy-Engel AM
  TITLE     Evolutionary conservation of the functional modularity of primate 
            and murine LINE-1 elements.
  JOURNAL   PLoS One. 2011 May 10;6(5):e19672. doi: 
            10.1371/journal.pone.0019672.
  PUBMED    21572950
REFERENCE   2  (bases 1 to 8430)
  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: "PLoS One.";
            date: "2011-05-10"; volume: "6(5):e19672. doi"; pages: " 
            10.1371/journal.pone.0019672"
COMMENT     Sequence Label: pBudORF2-CH
FEATURES             Location/Qualifiers
     source          1..8430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4554..4583
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4599..4616
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4647..4768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4658..4677)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4712..4731
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4898..5272)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(5291..5338)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5386..5702)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5400..5535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5454..5473)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5724..6886
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5955..6877
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6834..6854
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             6962..7003
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7013..7030
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7053..7070)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7059..7283
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(7415..8003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7495..7514)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8094..8112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8150..8172)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8272..8291
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg gatctgcagc aactaatgag caaaatcacc agctaacatc atagccgcca
      721 ccatgaccgg cagcaccagc cacatcacca tcctgaccct gaacatcaac ggcctgaaca
      781 gcgccatcaa gcggcaccgg ctggccagct ggatcaagag ccaggacccc agcgtgtgct
      841 gcatccagga gacccacctg acctgccggg acacccaccg gctgaagatt aagggctggc
      901 ggaagatcta ccaggccaac ggcaagcaga agaaggccgg cgtggccatc cttgtcagcg
      961 acaagaccga cttcaagccc accaagatca agcgggacaa ggagggccac tacatcatgg
     1021 tgaagggcag catccagcag gaggagctga ccatcctgaa catctacgcc cccaacactg
     1081 gtgccccccg gttcatcaaa caagttctga gcgaccttca gcgggacctg gacagccaca
     1141 ccctgatcat gggcgacttc aacacccccc tgagcaccct ggaccggagc acccggcaga
     1201 aggtgaacaa ggacacccag gagctgaaca gcgccctgca ccaggccgac ctgatcgaca
     1261 tctaccggac cctgcacccc aagagcaccg agtacacctt cttcagcgcc ccccaccaca
     1321 cctacagcaa gatcgaccac atcgtgggca gcaaggccct gctgagcaag tgcaagcgga
     1381 ctgagatcat caccaactac ctgagcgacc acagcgccat caagctggag ctgcggatca
     1441 agaacctgac ccagagccgg agcaccacct ggaagctgaa caacctgctg ctgaacgact
     1501 actgggttca caacgagatg aaggccgaga tcaagatgtt cttcgagacc aacgagaaca
     1561 aggacaccac ctaccagaac ctgtgggacg ccttcaaggc cgtgtgccgg ggcaagttca
     1621 tcgccctgaa cgcctacaag cggaagcagg agcggagcaa gatcgacacc ctgaccagcc
     1681 agctgaagga gctggagaag caggagcaga cccacagcaa ggccagtcgg cggcaggaga
     1741 tcaccaagat ccgggccgag ctgaaggaga tcgagaccca gaagaccctt cagaagatca
     1801 acgagagccg gagctggttc ttcgagcgga tcaacaagat cgaccggccc ctggcccggc
     1861 tgatcaagaa gaagcgggag aagaaccaga tcgacaccat caagaacgac aagggcgaca
     1921 tcaccaccga ccccaccgag atccagacca ccatccggga gtactacaag cacctgtacg
     1981 ccaacaagct ggagaatcta gaggagatgg acaccttcct ggacacctac accctgcccc
     2041 ggctgaacca ggaggaggtg gagagcctga accggcccat caccggcagc gagatcgtgg
     2101 ccatcatcaa cagcctgccc accaagaaga gccccggccc cgacggcttc accgccgagt
     2161 tctaccagcg gtacaaggag gagctggtgc ccttcctgct gaagctgttc cagagcatcg
     2221 agaaggaggg catcctgccc aacagcttct acgaggccag catcatcctg atccccaagc
     2281 ccggccggga caccaccaag aaggagaact tccggcccat cagcctgatg aacatcgacg
     2341 ccaagatcct gaacaagatc ctggccaacc ggattcagca gcacatcaag aagctgatcc
     2401 accacgacca agttggcttc atccccggca tgcaaggatg gttcaacatc cggaagagca
     2461 tcaacgtgat ccagcacatc aaccgggcca aggacaagaa ccacatgatc atcagcatcg
     2521 acgccgagaa ggctttcgac aagatccagc agcccttcat gctgaagacc ctgaacaagc
     2581 tgggcatcga cggcacctac ttcaagatca tccgggccat ctacgacaag cccaccgcca
     2641 acatcatcct gaacggccag aagctggagg ctttccccct gaagaccggc acccggcagg
     2701 gctgccccct gagccccctg ctgttcaaca tcgtgctgga ggtgctggcc cgggccatcc
     2761 ggcaggagaa ggagatcaag ggcatccagc tgggcaagga ggaggtgaag ctgagcctgt
     2821 tcgccgacga catgatcgtg tacctggaga accctattgt tagcgcccag aacctgctga
     2881 agctgatcag caacttcagt aaagttagcg gctacaagat caacgtgcag aagagccagg
     2941 ctttcctgta caccaataac cggcagaccg agagccagat catgggcgag ctgcccttca
     3001 ccatcgccag caagcggatc aagtacctgg gcatccagct gactcgggac gtcaaggacc
     3061 tgttcaagga gaactacaag cccctgctga aggagatcaa ggaggagacc aacaagtgga
     3121 agaacatccc ctgtagctgg gtgggccgga tcaacatcgt gaagatggcc atcctgccga
     3181 aagttatcta tcggttcaac gccatcccca tcaagctgcc catgaccttc ttcaccgagc
     3241 tggagaagac caccctgaag ttcatctgga accagaagcg ggcccggatc gccaagagca
     3301 tcctgagcca gaagaacaag gccggcggca tcaccctgcc cgacttcaag ctgtactaca
     3361 aggccaccgt gaccaagacc gcctggtact ggtaccagaa ccgggacatc gaccagtgga
     3421 accggaccga gcccagcgag atcatgcccc acatctacaa ctacctgatc ttcgacaagc
     3481 ccgagaagaa caagcagtgg ggcaaggaca gcctgttcaa caagtggtgc tgggagaact
     3541 ggctggccat ctgccggaag ctgaagctgg accccttcct gaccccctac accaagatca
     3601 acagccggtg gatcaaggac ctgaacgtga agcccaagac catcaagacc ctggaggaga
     3661 acctgggcat caccatccag gacatcggcg tgggcaagga cttcatgagc aagaccccca
     3721 aggccatggc caccaaggac aagatcgaca agtgggacct gatcaagctg aagagcttct
     3781 gcaccgccaa ggagaccacc atccgggtga accggcagcc caccacctgg gagaagattt
     3841 tcgccaccta cagcagcgac aagggcctga tcagccggat ctacaacgag ctgaagcaga
     3901 tttacaagaa gaagaccaac aaccccatca agaagtgggc caaggacatg aaccggcact
     3961 tcagcaagga ggacatctac gccgccaaga agcacatgaa gaagtgcagc agcagcctgg
     4021 ccatccggga gatgcagatc aagaccacca tgcggtatca cctgaccccc gtgcggatgg
     4081 ccatcatcaa gaagagcggc aacaaccgat gctggcgggg ctgcggcgag atcggcaccc
     4141 tgctgcactg ctggtgggac tgcaagctgg tgcagcccct gtggaagagc gtgtggcggt
     4201 tcctgcggga cctggagctg gagatcccct tcgaccccgc catccccctg ctgggcatct
     4261 accccaacga gtacaagagc tgctgctaca aggacacctg cacccggatg ttcatcgccg
     4321 ccctgttcac catcgccaag acctggaacc agcccaagtg ccccaccatg atcgactgga
     4381 tcaagaagat gtggcacatc tacactatgg agtactacgc cgccatcaag aacgacgagt
     4441 tcatcagctt cgtgggcacc tggatgaagc tggagaccat catcctgagc aagctgagcc
     4501 aggagcagaa gaccaagcac cggatcttca gcctgatcgg cggcaacgga tccgaacaaa
     4561 aactcatctc agaagaggat ctgaatatgc ataccggtca tcatcaccat caccattgag
     4621 tttgatcccc gggaattcag acatgataag atacattgat gagtttggac aaaccacaac
     4681 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4741 aaccattata agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc
     4801 cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat
     4861 agaaggcggc ggtggaatcg aaatctcgta gcacgtgtca gtcctgctcc tcggccacga
     4921 agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac ggctgctcgc
     4981 cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg acctccgacc
     5041 actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg ttgtccggca
     5101 ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc acaccggcga
     5161 agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac tcgaccgctc
     5221 cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttggcc atggtttagt
     5281 tcctcacctt gtcgtattat actatgccga tatactatgc cgatgattaa ttgtcaacac
     5341 gtgctgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     5401 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     5461 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     5521 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     5581 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     5641 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     5701 ccctcgtcga gctagcttcg tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc
     5761 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
     5821 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     5881 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     5941 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
     6001 atggcccttg cgtgccttga attacttcca cctggctcca gtacgtgatt cttgatcccg
     6061 agctggagcc aggggcgggc cttgcgcttt aggagcccct tcgcctcgtg cttgagttga
     6121 ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
     6181 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
     6241 tttctggcaa gatagtcttg taaatgcggg ccaggatctg cacactggta tttcggtttt
     6301 tgggcccgcg gccggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
     6361 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct
     6421 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
     6481 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctccag ggggctcaaa
     6541 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
     6601 ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca
     6661 cctcgattag ttctggagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
     6721 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
     6781 gatgtaattc tcgttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
     6841 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaacac gtggtcgcgg
     6901 ccgcttcgaa ggtaccagca cagtggactc gagagatctg gccggctggg cccgtttcga
     6961 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca
     7021 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     7081 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     7141 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     7201 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     7261 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagtggcg gtaatacggt
     7321 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7381 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7441 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7501 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7561 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7621 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7681 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7741 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7801 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7861 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7921 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7981 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8041 agtggaacga aaactcacgt taagggattt tggtcatgac attaacctat aaaaataggc
     8101 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     8161 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8221 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8281 agcagattgt actgagagtg caccatatat gcggtgtgaa ataccgcaca gatgcgtaag
     8341 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     8401 atcggtgcgg gcctcttcgc tattacgcca
//
