LOCUS       sequence-417614-        8580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An expression vector for mouse LINE1 ORF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8580)
  AUTHORS   Zheng F, Kawabe Y, Murakami M, Takahashi M, Nishihata K, Yoshida S, 
            Ito A, Kamihira M
  TITLE     LINE-1 vectors mediate recombinant antibody gene transfer by 
            retrotransposition in Chinese hamster ovary cells.
  JOURNAL   Biotechnol J. 2021 Jul;16(7):e2000620. doi: 10.1002/biot.202000620. 
            Epub 2021 May 20.
  PUBMED    33938150
REFERENCE   2  (bases 1 to 8580)
  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: "Biotechnol 
            J."; date: "2021-07"; volume: "16(7):e2000620. doi"; pages: " 
            10.1002/biot.202000620. Epub 2021 May 20"
COMMENT     Sequence Label: pORF2
FEATURES             Location/Qualifiers
     source          1..8580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..37)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        22..351
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      202..337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     401..420
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        401..419
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(4317..4891,4892..4894,4895..4903)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4484..4501)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4711..4730
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4904..5581
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(5034..5052)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(5084..5104)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     5302..5321
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5496..5519
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5625..5642)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5631..5855
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(5938..6526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6018..6037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6617..6635
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6673..6695)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6795..6814
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      6881..7336
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6968..6987)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7178..7199
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        7372..7675
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7676..7879
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7830..7850
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7876..7900
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        7997..8044
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8063..8437
                     /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"
     polyA_signal    join(8451..8580,1..5)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8488..8507)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8542..8561
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ggatcgatcc ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
       61 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
      121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
      181 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      241 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      301 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctctct
      361 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
      421 agacccaagc tggctagcgc taccggactc agatctcgac accatgcccc ccctgaccac
      481 caagatcacc ggcagcaaca actacttcag cctgatcagc ctgaacatca acggcctgaa
      541 cagccccatc aagcggcacc gcctgaccaa ctggctgcac aagcaggacc ccaccttctg
      601 ttgcctccag gagacccacc tgcgcgagaa ggaccggcac tacctgcgga tgaagggctg
      661 gaagaccatc tttcaggcca acggcatgaa gaagcaggct ggcgtggcca tcctgatcag
      721 cgacaagatc gacttccagc ccaaggtgat caagaaggac aaggagggcc acttcatcct
      781 gatcaagggc aagatcctgc aggaggagct gagcattctg aacatctacg cccccaacac
      841 ccgcgccgcc accttcacca aggagaccct cgtgaagctg aaggcccaca tcgctcccca
      901 caccatcatc gtcggcgact tcaacacccc cctgagcccc atggacagat cttggaagca
      961 gaagctgaac cgcgacaccc tgaagctgac cgaggtgatg aagcagatgg acctgaccga
     1021 catctaccgc accttctacc ccaagaccaa gggctacacc ttcttcagcg ctccccacgg
     1081 caccttcagc aagatcgacc acatcatcgg ccacaagagc gggctgaacc ggctgaagaa
     1141 catcgagatc gtgccctgca tcctgagcga ccaccatgcc ctgcgcctga tcttcaacaa
     1201 caagattaac aaccgcaagc ccaccttcac ctggaagctg aacaacaccc tgctgaacga
     1261 caccctggtg aaggaaggca tcaagaagga gatcaaggac ttcctggagt tcaacgagaa
     1321 cgaggccacc acctacccta acctgtggga caccatgaag gccttcctgc ggggcaagct
     1381 gatcgccatg agcgccttca agaagaagcg ggagcgcgcc cacactagta gcctgaccac
     1441 ccacctgaag gccctggaga agaaggaggc taactccccc aagcgctccc ggcggcagga
     1501 gatcatcaag ctgcgcggcg agatcaacca ggtggagacc cggcgcacca tccaacggat
     1561 caaccagacc cggtcttggt tcttcgagaa gatcaacaag atcgacaagc ccctggctcg
     1621 cctgaccaag ggccaccgcg acaagatcct gatcaacaag atccgcaacg agaagggcga
     1681 catcaccacc gaccccgagg agatccagaa caccatccgc agcttctaca agcgcctgta
     1741 cagcaccaag ctggagaacc tggacgagat ggacaagttc ctggaccgct atcaggtgcc
     1801 caagctgaac caggaccagg tggacctgct gaacagcccc atctccccca aggaaatcga
     1861 ggccgtgatc aacagcctgc ccgccaagaa gagccccggc cccgacggct tcagcgctga
     1921 gttctaccag accttcaagg aggacctgac ccccgtgctg cacaagcttt tccacaagat
     1981 cgaggtcgag ggcatcctcc ccaacagctt ctacgaggcc accatcaccc tgatccccaa
     2041 gccccagaag gaccccacca agatcgagaa cttccgcccc atcagcctga tgaacatcga
     2101 cgccaagatc ctgaacaaga tcctcgccaa ccgcatccag gagcacatca aggccatcat
     2161 ccaccctgac caggtcggct tcatccccgg catgcagggc tggttcaaca tccgcaagag
     2221 catcaacgtg atccactaca tcaacaagct gaaggacaag aaccacatga tcatcagcct
     2281 ggacgccgag aaggccttcg acaagatcca gcaccccttc atgatcaagg tcctggagcg
     2341 cagcggcatt cagggccagt acctcaacat gatcaaggcc atctacagca agcccgtcgc
     2401 caacatcaag gtgaacgggg agaagctgga ggccatccct ctgaagagcg gcacgcgtca
     2461 gggctgtcct ctgtccccct acctgttcaa catcgtgctg gaggtgctgg cccgcgctat
     2521 ccggcagcag aaggagatca agggcatcca gatcggcaag gaggaagtga agatcagcct
     2581 gttcgccgac gacatgatcg tgtacatcag cgaccccaag aacagcaacc gcgagctcct
     2641 gaacctgatc aacagcttcg gcgaggtggc tggctacaag atcaacagca acaagagcat
     2701 ggccttcctg tacaccaaga acaagcaggc cgagaaggag atccgcgaga ccaccccctt
     2761 cagcatcgcc accaacaaca tcaagtacct gggcgtgacc ctgaccaagg aggtgaagga
     2821 cctgtacgac aagaacttca agagcctgaa gaaggagatc aaggaggacc tgcgccgctg
     2881 gaaggacctg ccctgcagct ggatcggccg caccaacatc gtgaagatgg ccatcctgcc
     2941 caaggctatc taccgcttca acgccatccc catcaagatc cccacccagt tcttcaacga
     3001 gctcgagggc gccatctgca agttcatctg gaacaacaag aagccccgca tcgccaagac
     3061 cctgctgaag gacaagcgca ccagtggggg gatcaccatg cccgacctga agctgtacta
     3121 ccgcgccatc gtgatcaaga ccgcctggta ctggtaccgc gaccgccagg tggaccagtg
     3181 gaaccgcatc gaggaccccg agatgaaccc ccacacctac ggccacctga tcttcgacaa
     3241 gggcgccaag accatccagt ggaagaagga cagcatcttc aacaactggt gctggcacaa
     3301 ctggctgctg agctgccgcc gcatgcgcat cgacccctac ctaagcccct gcaccaaggt
     3361 gaagagcaag tggatcaagg agctgcacat caagcccgag actctgaagc tgatcgagga
     3421 gaaggtgggc aagagcctgg aggacatggg caccggcgaa aagttcctga accgcaccgc
     3481 catggcctgt gccgtgcgca gccggatcga taagtgggac ctgatgaagc tgcagtcctt
     3541 ctgtaaggcc aaggataccg tgtacaagac caagcgcccc ccgaccgact gggagcgcat
     3601 cttcacctac cccaagagcg accgcggcct gatcagcaac atctacaagg agctgaagaa
     3661 ggtggacctg cgcaagagca acaacccgct gaagaagtgg ggctccgagc tgaacaagga
     3721 gttctccccc gaggagtacc gcatggccga gaagcacctg aagaagtgca gcaccagcct
     3781 gatcatccgc gagatgcaga tcaagaccac cctgcgcttc cacctgaccc ccgtgcggat
     3841 ggccaagatc aagaacagcg gcgactcgcg atgctggcgg ggctgcggcg agcgcggcac
     3901 cctgctgcac tgctggtggg actgtcgcct ggtccagccc ctgtggaaga gcgtgtggcg
     3961 gttcctgcgg aagctggaca tcgtgctgcc cgaggacccc gctatccccc tgctgggcat
     4021 ctaccctgag gaggccccca ccggcaagaa ggacacctgc agtaccatgt tcatcgccgc
     4081 tctgttcatc atcgcccgca actggaagga gccccgctgc cccagcaccg aggagtggat
     4141 tcagaagatg tggtacatct acaccatgga gtactacagc gccatcaaga agaacgagtt
     4201 catgaagttc ctggccaagt ggatggacct ggagagcatc atcctgagcg aggtgaccca
     4261 gagccagcgc aacagccaca acatgtacag cctgatcagc ggctactagg gatccgcccc
     4321 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     4381 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     4441 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     4501 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     4561 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     4621 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     4681 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     4741 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtacaca
     4801 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     4861 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggcct cctccgagga
     4921 cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     4981 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     5041 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcca
     5101 gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca agaagctgtc
     5161 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     5221 cgtgacccag gactcctccc tgcaggacgg ctccttcatc tacaaggtga agttcatcgg
     5281 cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct gggaggcctc
     5341 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca aggccctgaa
     5401 gctgaaggac ggcggccact acctggtgga gttcaagtct atctatatgg ccaagaagcc
     5461 cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct cccacaacga
     5521 ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc tgttcctgta
     5581 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     5641 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5701 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5761 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5821 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagcat
     5881 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5941 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6001 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6061 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6121 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6181 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6241 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6301 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6361 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6421 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6481 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6541 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6601 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     6661 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6721 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6781 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     6841 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg acgcgccctg tagcggcgca
     6901 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6961 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     7021 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     7081 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     7141 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     7201 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     7261 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     7321 ttaacgctta caatttccat tcgccattca ggctgaacta gatctagagt ccgttacata
     7381 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7441 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7501 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7561 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7621 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     7681 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     7741 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     7801 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     7861 ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
     7921 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc gggaacggtg
     7981 cattggaacg gaccgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     8041 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     8101 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     8161 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     8221 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     8281 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     8341 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     8401 caactgcgtg cacttcgtgg ccgaggagca ggactgacac tcgacctcga aacttgttta
     8461 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     8521 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
//
