LOCUS       sequence-78373-e        5582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the codon optimized ORF1 of L1RP with his tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5582)
  AUTHORS   Wagstaff BJ, Kroutter EN, Derbes RS, Belancio VP, Roy-Engel AM
  TITLE     Molecular reconstruction of extinct LINE-1 elements and their 
            interaction with nonautonomous elements.
  JOURNAL   Mol Biol Evol. 2013 Jan;30(1):88-99. doi: 10.1093/molbev/mss202. 
            Epub 2012 Aug 23.
  PUBMED    22918960
REFERENCE   2  (bases 1 to 5582)
  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: 
            "10.1093/molbev/mss202"; journalName: "Mol Biol Evol"; date: 
            "2013-01"; volume: "30"; issue: "1"; pages: "88-99"
COMMENT     Sequence Label: pBudORF1-CH
FEATURES             Location/Qualifiers
     source          1..5582
                     /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             1706..1735
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1751..1768
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1799..1920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1810..1829)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1864..1883
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2050..2424)
                     /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(2443..2490)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2538..2854)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2552..2687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2606..2625)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2876..4038
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3107..4029
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3986..4006
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4114..4155
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4165..4182
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4205..4222)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4211..4435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4567..5155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4647..4666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5246..5264
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5302..5324)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5424..5443
                     /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 cccaagctta gctaaaacca caaagatggg caagaagcag aaccgcaaga ccggcaacag
      721 caagacccag agcgccagcc ccccccccaa ggagcgcagc agcagccccg ccaccgagca
      781 gagctggatg gagaacgact tcgacgagct gcgcgaggag ggcttccgcc gcagcaacta
      841 cagcgagctg cgcgaggaca tccagaccaa gggcaaggag gtggagaact tcgagaagaa
      901 cctggaggag tgcatcaccc gcatcaccaa caccgagaag tgcctgaagg agctgatgga
      961 gctgaagacc aaggcccgcg agctgcgcga ggagtgccgc agcctgcgca gccgctgcga
     1021 ccagctggag gagcgcgtga gcgcaatgga ggacgagatg aacgagatga agcgcgaggg
     1081 caagttccgc gagaagcgca tcaagcgcaa cgagcagagc ctccaggaga tatgggacta
     1141 cgtgaagcgc cccaacctgc gcctgatcgg cgtgcccgag agcgacgtgg agaacggcac
     1201 caagctggag aacaccctac aggacatcat ccaggagaac ttccccaacc tggcccgcca
     1261 ggccaacgtg cagatccagg agatccagcg caccccccag cgctacagca gccgccgcgc
     1321 caccccccgc cacatcatcg tgcgcttcac caaggtggag atgaaggaga agatgctgcg
     1381 cgccgcccgc gagaagggcc gggttaccct gaagggcaag cccatccgcc tgaccgccga
     1441 cctgagcgcc gagaccctac aggcccgccg cgagtggggg ccaatcttca acatcctgaa
     1501 ggagaagaac ttccagcccc gcatcagcta ccccgccaag ctgagcttca tcagcgaggg
     1561 cgagatcaaa tacttcatcg acaagcagat gctgcgcgac ttcgtgacca cccgccccgc
     1621 cctgaaggag ctgctgaagg aggccctgaa catggagcgc aacaaccggt atcagcccct
     1681 gcaaaaccac gccaagatgg gatccgaaca aaaactcatc tcagaagagg atctgaatat
     1741 gcataccggt catcatcacc atcaccattg agtttgatcc ccgggaattc agacatgata
     1801 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1861 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1921 ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
     1981 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
     2041 tagcacgtgt cagtcctgct cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg
     2101 cagggcgaac tcccgccccc acggctgctc gccgatctcg gtcatggccg gcccggaggc
     2161 gtcccggaag ttcgtggaca cgacctccga ccactcggcg tacagctcgt ccaggccgcg
     2221 cacccacacc caggccaggg tgttgtccgg caccacctgg tcctggaccg cgctgatgaa
     2281 cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa
     2341 cccgagccgg tcggtccaga actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg
     2401 aacggcactg gtcaacttgg ccatggttta gttcctcacc ttgtcgtatt atactatgcc
     2461 gatatactat gccgatgatt aattgtcaac acgtgctgat cagatccgaa aatggatata
     2521 caagctcccg ggagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     2581 tggaatagct cagaggcaga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     2641 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     2701 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     2761 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     2821 ctgcctgctg gggagcctgg ggactttcca caccctcgtc gagctagctt cgtgaggctc
     2881 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     2941 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     3001 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     3061 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
     3121 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
     3181 cacctggctc cagtacgtga ttcttgatcc cgagctggag ccaggggcgg gccttgcgct
     3241 ttaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
     3301 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     3361 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     3421 ggccaggatc tgcacactgg tatttcggtt tttgggcccg cggccggcga cggggcccgt
     3481 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     3541 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     3601 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     3661 ccgcttcccg gccctgctcc agggggctca aaatggagga cgcggcgctc gggagagcgg
     3721 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     3781 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctggag cttttggagt
     3841 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     3901 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctcgttgga atttgccctt
     3961 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     4021 ccatttcagg tgtcgtgaac acgtggtcgc ggccgcttcg aaggtaccag cacagtggac
     4081 tcgagagatc tggccggctg ggcccgtttc gaaggtaagc ctatccctaa ccctctcctc
     4141 ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga
     4201 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4261 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4321 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4381 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4441 gaggcggaaa gaaccagtgg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4501 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4561 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4621 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4681 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4741 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4801 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4861 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4921 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4981 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5041 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5101 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5161 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5221 tttggtcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5281 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     5341 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     5401 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     5461 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
     5521 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     5581 ca
//
