LOCUS       sequence-422244-        7481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full length human LINE-1 ORF2p Core (1-1275) with a 
            C-terminal 3xFlag tag for baculovirus production in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7481)
  AUTHORS   Baldwin ET, van Eeuwen T, Hoyos D, Zalevsky A, Tchesnokov EP, 
            Sanchez R, Miller BD, Di Stefano LH, Ruiz FX, Hancock M, Isik E, 
            Mendez-Dorantes C, Walpole T, Nichols C, Wan P, Riento K, Halls-Kass
            R, Augustin M, Lammens A, Jestel A, Upla P, Xibinaku K, Congreve S, 
            Hennink M, Rogala KB, Schneider AM, Fairman JE, Christensen SM, 
            Desrosiers B, Bisacchi GS, Saunders OL, Hafeez N, Miao W, Kapeller 
            R, Zaller DM, Sali A, Weichenrieder O, Burns KH, Gotte M, Rout MP, 
            Arnold E, Greenbaum BD, Romero DL, LaCava J, Taylor MS
  TITLE     Structures, functions and adaptations of the human LINE-1 ORF2 
            protein.
  JOURNAL   Nature. 2024 Feb;626(7997):194-206. doi: 10.1038/s41586-023-06947-z.
            Epub 2023 Dec 14.
  PUBMED    38096902
REFERENCE   2  (bases 1 to 7481)
  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: 
            "10.1038/s41586-023-06947-z"; journalName: "Nature"; date: 
            "2024-02"; volume: "626"; issue: "7997"; pages: "194-206"
COMMENT     Sequence Label: pMT692 ORF2p-3C-3xF (1-1275, ORFeus-Hs) in
            pDA...
FEATURES             Location/Qualifiers
     source          1..7481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  1..224
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     rep_origin      complement(288..876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(368..387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(915..1706,1707..1775))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1538..1557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1776..1847)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  1872..2037
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        2202..2293
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2218..2235
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             6172..6195
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6196..6261
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        join(6665..6670,6671..6687,6688..6693)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6882..7415
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gaatatagtc tttacatgcc cttggccacg cccctcttta atacgacggg caatttgcac
       61 ttcagaaaat gaagagtttg ctttagccat aacaaaagtc cagtatgctt tttcacagca
      121 taactggact gatttcagtt tacaactatt ctgtctagtt taagacttta ttgtcatagt
      181 ttagatctat tttgttcagt ttaagacttt attgtccgcc cacacccgct tacgcagggc
      241 atctgcgcat ccacaggaag ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      481 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      721 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      901 gggtctgacg cttattacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
      961 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     1021 ttaccatctg gccccagtgc tgcaatgata ccgcgtgacc cacgctcacc ggctccagat
     1081 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     1141 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     1201 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     1261 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1321 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     1381 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1441 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1501 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     1561 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1621 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1681 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1741 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1801 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttaggaccca
     1861 agtcaaccgg ttgtgggcgg acaaaatagt tgggaactgg gaggggtgga aatggagttt
     1921 ttaaggatta tttagggaag agtgacaaaa tagatgggaa ctgggtgtag cgtcgtaagc
     1981 taatacgaaa attaaaaatg acaaaatagt ttggaactag atttcactta tctggttatt
     2041 taaattaggg ataacagggt aatagtgcga gaccggtctc gcaggtaggg ataacagggt
     2101 aatcagcctg cggtccgggc atagtacgca gcttcttcta gttcaattac accatttttt
     2161 agcagcaccg gattaacata actttccaaa atgttgtaga tatcatggag ataattaaaa
     2221 tgataaccat ctcgcaaata aataagtatt ttactgtttt cgtaacagtt ttgtaataaa
     2281 aaaacctata aatagttggc caggcctagt gcggccgcca ccatgaccgg cagcaacagc
     2341 cacatcacca tcctgaccct gaacatcaac ggcctgaaca gcgccatcaa gcgccaccgc
     2401 ctggccagct ggatcaagag ccaggacccc agcgtgtgct gcatccagga gacccacctg
     2461 acctgccgcg acacccaccg cctgaagatc aagggctggc gcaagatcta ccaggccaac
     2521 ggcaagcaga agaaggccgg cgtggccatc ctggtgagcg acaagaccga cttcaagccc
     2581 accaagatca agcgcgacaa ggagggccac tacatcatgg tgaagggcag catccagcag
     2641 gaggagctga ccatcctgaa catctacgcc cccaacaccg gtgccccccg cttcatcaag
     2701 caggtgctga gcgacctgca gcgcgacctg gacagccaca ccctgatcat gggcgacttc
     2761 aacacccccc tgagcaccct ggaccgcagc acccgccaga aggtgaacaa ggacacccag
     2821 gagctgaaca gcgccctgca ccaggccgac ctgatcgaca tctaccgcac cctgcacccc
     2881 aagagcaccg agtacacctt cttcagcgcc ccccaccaca cctacagcaa gatcgaccac
     2941 atcgtgggca gcaaggccct gctgagcaag tgcaagcgca ccgagatcat caccaactac
     3001 ctgagcgacc acagcgccat caagctggag ctgcgcatca agaacctgac ccagagccgc
     3061 agcaccacct ggaagctgaa caacctgctg ctgaacgact actgggtgca caacgagatg
     3121 aaggccgaga tcaagatgtt cttcgaaacc aacgagaaca aggacaccac ctaccagaac
     3181 ctgtgggacg ccttcaaggc cgtgtgccgc ggcaagttca tcgccctgaa cgcctacaag
     3241 cgcaagcagg agcgcagcaa gatcgacacc ctgaccagcc agctgaagga gctggagaag
     3301 caggagcaga cccacagcaa ggccagccgc cgccaggaga tcaccaagat ccgcgccgag
     3361 ctgaaggaga tcgagaccca gaagaccctg cagaagatca acgagtcgcg aagctggttc
     3421 ttcgagcgca tcaacaagat cgaccgcccc ctggcccgcc tgatcaagaa gaagcgcgag
     3481 aagaaccaga tcgacaccat caagaacgac aagggcgaca tcaccaccga ccccaccgag
     3541 atccagacca ccatccgcga gtactacaag cacctgtacg ccaacaagct ggagaacctg
     3601 gaggagatgg acaccttcct ggacacctac accctgcccc gcctgaacca ggaggaggtg
     3661 gagagcctga accgccccat caccggcagc gagatcgtgg ccatcatcaa cagcctgccc
     3721 accaagaaga gccccggccc cgacggcttc accgccgagt tctaccagcg ctacaaggag
     3781 gagctggtgc ccttcctgct gaagctgttc caaagcatcg agaaggaggg catcctgccc
     3841 aacagcttct acgaggccag catcatcctg atccccaagc ccggccgcga caccaccaag
     3901 aaggagaact tccgccccat cagcctgatg aacatcgacg ccaagatcct gaacaagatc
     3961 ctggccaacc gcatccagca gcacatcaag aagctgatcc accacgacca ggtgggcttc
     4021 atccccggga tgcagggctg gttcaacatc cgcaagagca tcaacgtgat ccagcacatc
     4081 aaccgcgcca aggacaagaa ccacatgatc atcagcatcg acgccgagaa ggccttcgac
     4141 aagatccagc agcccttcat gctgaagacc ctgaacaagc tgggcatcga cggcacctac
     4201 ttcaagatca tccgcgccat ctacgacaag cccaccgcca acatcatcct gaacggccag
     4261 aagctggagg ccttccccct gaagaccggc acgcgtcagg gctgccccct gagccccctg
     4321 ctgttcaaca tcgtgctgga ggtgctggcc cgcgccatcc gccaggagaa ggagatcaag
     4381 ggcatccagc tgggcaagga ggaggtgaag ctgagcctgt tcgccgacga catgatcgtg
     4441 tacctggaga accccatcgt gagcgcccag aacctgctga agctgatcag caacttcagc
     4501 aaggtgagcg gctacaagat caacgtgcag aagagccagg ccttcctgta caccaacaac
     4561 cgccagaccg agagccagat catgggcgag ctgcccttca ccatcgctag caagcgcatc
     4621 aagtacctgg gcatccagct gacccgcgac gtgaaggacc tgttcaagga gaactacaag
     4681 cccctgctga aggagatcaa ggaggagacc aacaagtgga agaacatccc ctgcagctgg
     4741 gtgggccgca tcaacatcgt gaagatggcc atcctgccca aggtgatcta ccgcttcaac
     4801 gccatcccca tcaagctgcc catgaccttc ttcaccgagc tggagaagac caccctgaag
     4861 ttcatctgga accagaagcg cgcccgcatc gccaagagca tcctgagcca gaagaacaag
     4921 gccggcggca tcaccctgcc cgacttcaag ctgtactaca aggccaccgt gaccaagacc
     4981 gcctggtact ggtaccagaa ccgcgatatc gaccagtgga accgcaccga gcccagcgag
     5041 atcatgcccc acatctacaa ctacctgatc ttcgacaagc ccgagaagaa caagcagtgg
     5101 ggcaaggaca gcctgttcaa caagtggtgc tgggagaact ggctggccat ctgccgcaag
     5161 ctgaagctgg accccttcct gaccccctac accaagatca acagccgctg gatcaaggac
     5221 ctgaacgtga agcccaagac catcaagacc ctggaggaga acctgggcat caccatccag
     5281 gacatcggcg tgggcaagga cttcatgagc aagaccccca aggccatggc caccaaggac
     5341 aagatcgaca agtgggacct gatcaagctg aagagcttct gcaccgccaa ggagaccacc
     5401 atccgcgtga accgccagcc caccacctgg gagaagatct tcgccaccta cagcagcgac
     5461 aagggcctga tcagccgcat ctacaacgag ctgaagcaga tctacaagaa gaagaccaac
     5521 aaccccatca agaagtgggc caaggacatg aaccgccact tcagcaagga ggacatctac
     5581 gccgccaaga agcatatgaa gaagtgcagc agcagcctgg ccatccgcga gatgcagatc
     5641 aagaccacca tgcgctacca cctgaccccc gtgcgcatgg ccatcatcaa gaagagcggc
     5701 aacaaccgct gctggcgcgg ctgcggcgag atcggcaccc tgctgcactg ctggtgggac
     5761 tgcaagctgg tgcagcccct gtggaagagc gtgtggcgct tcctgcgcga cctggagctg
     5821 gagatcccct tcgaccccgc catccccctg ctgggcatct accccaacga gtacaagagc
     5881 tgctgctaca aggacacctg cacccgcatg ttcatcgccg ccctgttcac catcgccaag
     5941 acctggaacc agcccaagtg ccccaccatg atcgattgga tcaagaagat gtggcacatc
     6001 tacaccatgg agtactacgc cgccatcaag aacgacgagt tcatcagctt cgtgggcacc
     6061 tggatgaagc tggagaccat catcctgagc aagctgagcc aggagcagaa gaccaagcac
     6121 cgcatcttca gcctgatcgg cggcaacggc ggcggcagcg gcggcggcag cctggaagtt
     6181 ctgttccagg ggcccgacta caaagaccat gacggtgatt ataaagatca tgacatcgat
     6241 tacaaggatg acgatgacaa gtagtgataa tggcctgtgg ccgtaacttg tagcagcaat
     6301 ctagctttgt cgatattcgt ttgtgttttg ttttgtaata aaggttcgac gtcgttcaaa
     6361 atattatgcg cttttgtatt tctttcatca ctgtcgttag tgtacaattg actcgacgta
     6421 aacacgttaa ataaagcttg gacatattta acatcgggcg tgttagcttt attaggccga
     6481 ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt ccgagaacga ttttgccata
     6541 gccacacgac gcctattaat tgtgtcggct aacacgtccg cgatcaaatt tgtagttgag
     6601 ctttttggaa ttatttctga ttgcgggcgt tttttcgctg ggacgcccga acggtggcta
     6661 ctcgttgaca taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg
     6721 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     6781 ggcttgttat gactgttttt ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     6841 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagcaacg
     6901 atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca
     6961 ttcgtacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt
     7021 tcggtcgtga gttcggtgat gtagccacct actcccaaca tcagccggac tccgattacc
     7081 tcgggaactt gctccgtagt aagacattta ttgcgcttgc tgccttcgac caagaagcgg
     7141 ttgttggcgc tctcgcggct tacgttctgc ccaagtttga gcagccgcgt agtgagatct
     7201 atatctatga tctcgcagtc tccggcgagc accggaggca gggcattgcc accgcgctca
     7261 tcaatctcct caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag
     7321 attacggtga cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga
     7381 tgcactttga tatcgaccca agtaccgcca cctaacaatt cgttcaagcc gagatcggct
     7441 tcccggccgc ggagttgttc ggtaaattgt cacaacgccg c
//
