LOCUS       sequence-289085-        6875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsPatL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6875)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6875)
  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"
COMMENT     Sequence Label: pCMV-TAPtagN-HsPatL1_J
FEATURES             Location/Qualifiers
     source          1..6875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      658..667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..867
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             868..1041
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1078..1098
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1105..1182
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    3663..3784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3700..3719)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3754..3773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3791..4246)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3928..3949)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4140..4159
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4273..4377
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4379..4736
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4379..4570
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4402..4422)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4587..4722
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4771..5565
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4771..5565
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4825..4844)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5435..5454
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5753..5772)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5797..5844
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6173..6761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6662..6681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtgcc
      661 accatggcag gccttgcgca acacgatgaa gccgtggaca acaaattcaa caaagaacaa
      721 caaaacgcgt tctatgagat cttacattta cctaacttaa acgaagaaca acgaaacgcc
      781 ttcatccaaa gtttaaaaga tgacccaagc caaagcgcta accttttagc agaagctaaa
      841 aagctaaatg atgctcaggc gccgaaagta gacaacaaat tcaacaaaga acaacaaaac
      901 gcgttctatg agatcttaca tttacctaac ttaaacgaag aacaacgaaa cgccttcatc
      961 caaagtttaa aagatgaccc aagccaaagc gctaaccttt tagcagaagc taaaaagcta
     1021 aatgatgctc aggcgccgaa agtagacgcg aattgtgata tacctacaac tgcttctgaa
     1081 aatttatatt ttcaaggtga actaaagaga agatggaaaa agaatttcat agccgtctca
     1141 gcagccaacc gctttaagaa aatctcatcc tccggggcac ttgatccatc cggactcaga
     1201 tctcgagcta tgttccgcta cgagtctttg gaggattgtc ctctggatga agatgaagat
     1261 gcatttcagg gactgggaga agaagatgaa gagattgatc aattcaatga tgatacattt
     1321 gggtcaggtg cagttgatga tgattggcag gaagcacatg agcgcctggc tgaattggaa
     1381 gaaaagctac cagtggcagt taatgaacaa acaggcaatg gagagagaga tgaaatggac
     1441 ttgttgggtg accatgagga gaatctggca gaaaggctca gtaagatggt gattgaaaat
     1501 gaactagaag atccagctat tatgagggca gtgcagacca ggccagtttt acaaccccaa
     1561 ccaggaagtc tgaattccag tatctgggat ggatctgaag ttctgaggcg aatccgagga
     1621 ccactgcttg ctcaggaaat gcctacagtg tctgtattag aatatgcttt gcctcagagg
     1681 cccccccagg gtccagaaga tgatcgggac ctttctgaac gagcattacc aaggcggtca
     1741 acttcaccta tcattggcag tcctcctgtt agagctgtcc ccataggcac cccacctaaa
     1801 cagatggctg tacccagctt cacccaacag attctgtgtc cgaagcctgt ccatgttcgg
     1861 cccccaatgc cacctcgtta tcctgctccc tatggtgaga ggatgtctcc aaaccagctc
     1921 tgcagtgtcc cgaactcttc cctcctgggt cacccttttc ctcctagtgt tcctcctgtt
     1981 ctcagccccc tccagagagc acagcttctt ggaggagcac agctacagcc tggacggatg
     2041 tctcccagcc agtttgcacg ggtccctgga tttgttggta gtccacttgc tgccatgaat
     2101 cccaagttgc tacaagggcg agttgggcag atgcttcccc cagcaccagg cttccgtgcc
     2161 ttctttagtg ctccaccctc cgctacacca cctccacagc agcaccctcc tggcccagga
     2221 ccccacctgc aaaacctaag atctcaggcc ccaatgttta gaccggacac aactcacctc
     2281 catccacagc accgtcgact cttgcatcag agacagcaac agaatagaag tcagcatcgg
     2341 aatctcaatg gtgcgggaga tagaggaagt caccggagca gtcatcaaga tcatctccga
     2401 aaggatccat atgccaatct catgttgcag cgggaaaagg attgggtctc taaaatccag
     2461 atgatgcaac tgcaaagcac tgatccctac ctggatgatt tttattacca gaattacttt
     2521 gaaaaactgg agaaactgtc agctgctgaa gaaatacaag gtgatggccc taagaaggag
     2581 cgcaccaagc ttatcacccc tcaggtggcc aaactggagc acgcctataa gccagtgcaa
     2641 tttgagggct ctttgggaaa gcttaccgtt tctagtgtga ataatccccg aaaaatgatt
     2701 gatgctgttg tgacatctcg gagtgaggat gatgagacaa aagaaaaaca agttcgagac
     2761 aagaggagaa aaacccttgt tataattgag aaaacctaca gcttactcct tgatgtggag
     2821 gactatgaaa gacgttatct cctaagtctg gaagaagagc gacctgccct aatggatgac
     2881 agaaagcaca aaatttgtag catgtatgac aacttaaggg ggaaattgcc tggacaagag
     2941 aggcctagtg atgaccactt tgtacagatc atgtgtatcc gaaaagggaa gagaatggtt
     3001 gcccgtattc ttcctttcct ctccacagag caagcagctg acattctcat gacaacagcc
     3061 aggaacctcc ctttccttat caagaaggat gcacaagatg aggtgctgcc atgcttactg
     3121 agtcccttct ctctccttct ctatcatctt ccatcagtga gtatcaccag ccttttgcga
     3181 cagctaatga acctacctca aagtgcagct acaccagcac tctccaatcc tcacctcact
     3241 gctgtgctcc agaacaagtt tggcctgtca ctgctcctca tcctcctgag ccgtggtgaa
     3301 gacctacaga gttcagaccc tgctacagaa tcaacacaaa ataatcagtg gacggaggtg
     3361 atgttcatgg caacacgaga acttctgcgg attccccaag cagccctggc caagccaatc
     3421 tctataccta caaacctagt gtccctcttt tctcgctatg ttgaccggca gaaactgaac
     3481 ttgctggaga caaaactgca gctagttcag gggatacgat aaccgcgggc ccgggatcca
     3541 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3601 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3661 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3721 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3781 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3841 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3901 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3961 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4021 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4081 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4141 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4201 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4261 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4321 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4381 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4441 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4501 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4561 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4621 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4681 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4741 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4801 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4861 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4921 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4981 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5041 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5101 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5161 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5221 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5281 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5341 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5401 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5461 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5521 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5581 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5641 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5701 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5761 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5821 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5881 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5941 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     6001 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     6061 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6121 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6181 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6241 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6301 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6361 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6421 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6481 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6541 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6601 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6661 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6721 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6781 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6841 ctgattctgt ggataaccgt attaccgcca tgcat
//
