LOCUS       sequence-417609-        5828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An expression vector for mouse LINE1 ORF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5828)
  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 5828)
  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: pORF1
FEATURES             Location/Qualifiers
     source          1..5828
                     /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(1565..2139,2140..2142,2143..2151)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1732..1749)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1959..1978
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2152..2829
                     /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(2282..2300)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2332..2352)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2550..2569
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2744..2767
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2873..2890)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2879..3103
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3186..3774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3266..3285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3865..3883
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3921..3943)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4043..4062
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4129..4584
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4216..4235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4426..4447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        4620..4923
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4924..5127
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5078..5098
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5124..5148
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        5245..5292
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5311..5685
                     /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(5699..5828,1..5)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5736..5755)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5790..5809
                     /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 tggctagcac catggccaag ggcaagcgcc gcaacctgac caaccgcaac
      481 caggaccaca gccccagccc cgagcccagc acccccacca gccccagccc cggcaacccc
      541 aacacccccg agaacctgga cctggacctg aaggcctacc tgatgatgat ggtggaggac
      601 atcaagaagg acttcaacaa gagcctgaag gagatccagg agaacaccgc caaggagctg
      661 caggtgctga aggagaagca ggagaacacc atcaagcagg tggaggtgct gaccgagaag
      721 gaggagaaga cctacaagca ggtgatggag atgaacaaga ccatcctgga cctgaagcgc
      781 gaggtcgaca ccatcaagaa gacccagagc gaggccaccc tggagatcga gaccctgggc
      841 aagaagagcg gcaccatcga cctgagcatc agcaaccgca tccaggagat ggaggagcgc
      901 atcagcggcg ccgaggacag catcgagaac atcggcacca ccatcaagga gaacggcaag
      961 tgcaagaaga tcctgaccca gaacatccag gagatccagg acaccatccg ccgccccaac
     1021 gtgcgcatca tcggcgtgga cgagaacgag gacttccagc tgaagggccc cgccaacatc
     1081 ttcaacaaga tcatcgagga gaacttcccc aacctgaaga acgagatgca catgaacatc
     1141 caggaggcct accgcacccc caaccgcctg gaccagaagc gcaacagctc tagacacatc
     1201 atcatccgca ccagcaacgc cctgaacaag gaccgcatcc tgaaggccgt gcgcgagaag
     1261 ggccaggtga cctacaaggg caagcccatc cgcatcaccc ccgacttcag ccccgagacc
     1321 atgaaggccc gccgcgcctg gaccgacgtg atccagaccc tgcgcgagca caagctgcag
     1381 ccccgcctgc tgtaccccgc caagctgagc atcatcatcg agggcgagac caaggtgttc
     1441 cacgacaaga ccaagttcac ccactacctg agcaccaacc ccgccctgca gcgcatcatc
     1501 accgagaaga accagtacaa gaacggcaac aacgccctgg agaagacccg ccgctaagga
     1561 tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     1621 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     1681 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     1741 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     1801 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     1861 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     1921 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     1981 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     2041 ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     2101 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac catggcctcc
     2161 tccgaggacg tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac
     2221 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2281 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
     2341 cagttccagt acggctccaa ggtgtacgtg aagcaccccg ccgacatccc cgactacaag
     2401 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     2461 gtggtgaccg tgacccagga ctcctccctg caggacggct ccttcatcta caaggtgaag
     2521 ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc agaagaagac tatgggctgg
     2581 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccacaag
     2641 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagtctat ctatatggcc
     2701 aagaagcccg tgcagctgcc cggctactac tacgtggact ccaagctgga catcacctcc
     2761 cacaacgagg actacaccat cgtggagcag tacgagcgcg ccgagggccg ccaccacctg
     2821 ttcctgtagc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2881 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2941 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3001 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3061 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3121 accagcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3601 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3841 ttggtcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     3901 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3961 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4021 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     4081 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggac gcgccctgta
     4141 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4201 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4261 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4321 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4381 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4441 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4501 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4561 acaaaatatt aacgcttaca atttccattc gccattcagg ctgaactaga tctagagtcc
     4621 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     4681 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     4741 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     4801 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     4861 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     4921 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     4981 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     5041 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     5101 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
     5161 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggccgg
     5221 gaacggtgca ttggaacgga ccgtgttgac aattaatcat cggcatagta tatcggcata
     5281 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     5341 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     5401 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     5461 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     5521 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     5581 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     5641 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacactc gacctcgaaa
     5701 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5761 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5821 tcatgtct
//
