LOCUS       sequence-393221-        6203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid encoding the first open reading frame 
            (ORF1, expressing the protein ORF1p) of human L1 (L1.3); carries a 
            C-terminus Flag tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6203)
  AUTHORS   Cook PR, Jones CE, Furano AV
  TITLE     Phosphorylation of ORF1p is required for L1 retrotransposition.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4298-303. doi: 
            10.1073/pnas.1416869112. Epub 2015 Mar 23.
  PUBMED    25831499
REFERENCE   2  (bases 1 to 6203)
  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: 
            "10.1073/pnas.1416869112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-04-7- 7"; volume: "112"; issue: "14"; pages: "4298-303"
COMMENT     Sequence Label: pORF1-Flag
FEATURES             Location/Qualifiers
     source          1..6203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1952..1975
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2049..2066)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2055..2279
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2325..2753
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2412..2431)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2622..2643
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2762..2782)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2767..3096
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2947..3082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3009..3028
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3117..3716
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3117..3136)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3613..3633
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3876..3997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3913..3932)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3967..3986
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4046..4062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4046..4062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4059..4081)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4070..4086
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4094..4100,4101..4118,4119..4124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4139..4160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4277..4294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4448..5036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4528..4547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5207..5998,5999..6067))
                     /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     5830..5849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6068..6172)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgcaatg gggaaaaaac agaacagaaa
      961 aactggaaac tctaaaacgc agagcgcctc tcctcctcca aaggaacgca gttcctcacc
     1021 agcaacagaa caaagctgga tggagaatga ttttgacgag ctgagagaag aaggcttcag
     1081 acgatcaaat tactctgagc tacgggagga cattcaaacc aaaggcaaag aagttgaaaa
     1141 ctttgaaaaa aatttagaag aatgtataac tagaataacc aatacagaga agtgcttaaa
     1201 ggagctgatg gagctgaaaa ccaaggctcg agaactacgt gaagaatgca gaagcctcag
     1261 gagccgatgc gatcaactgg aagaaagggt atcagcaatg gaagatgaaa tgaatgaaat
     1321 gaagcgagaa gggaagttta gagaaaaaag aataaaaaga aatgagcaaa gcctccaaga
     1381 aatatgggac tatgtgaaaa gaccaaatct acgtctgatt ggtgtacctg aaagtgatgt
     1441 ggagaatgga accaagttgg aaaacactct gcaggatatt atccaggaga acttccccaa
     1501 tctagcaagg caggccaacg ttcagattca ggaaatacag agaacgccac aaagatactc
     1561 ctcgagaaga gcaactccaa gacacataat tgtcagattc accaaagttg aaatgaagga
     1621 aaaaatgtta agggcagcca gagagaaagg tcgggttacc ctcaaaggaa agcccatcag
     1681 actaacagtg gatctctcgg cagaaaccct acaagccaga agagagtggg ggccaatatt
     1741 caacattctt aaagaaaaga attttcaacc cagaatttca tatccagcca aactaagctt
     1801 cataagtgaa ggagaaataa aatactttat agacaagcaa atgttgagag attttgtcac
     1861 caccaggcct gccctaaaag agctcctgaa ggaagcacta aacatggaaa ggaacaaccg
     1921 gtaccagccg ctgcaaaatc atgccaaaat ggattataag gacgacgacg acaagtagga
     1981 attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg
     2041 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2101 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2161 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2221 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2281 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     2341 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2401 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2461 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2521 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2581 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2641 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2701 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     2761 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2821 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2881 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctcact
     2941 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3001 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3061 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccggggcc gccaccatga
     3121 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     3181 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     3241 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     3301 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     3361 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     3421 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     3481 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     3541 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     3601 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     3661 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgattcg
     3721 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     3781 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3841 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     3901 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3961 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4021 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4081 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4141 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4201 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4261 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4321 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4381 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4861 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5581 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6121 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6181 cccgaaaagt gccacctgac gtc
//
