LOCUS       sequence-421013-        6232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmaug2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6232)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6232)
  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: pLNHA-C1-HsSmaug2_Q
FEATURES             Location/Qualifiers
     source          1..6232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..30)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      195..330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             379..1173
                     /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             379..1173
                     /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(433..452)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1043..1062
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1361..1380)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1405..1452
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1781..2369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2270..2289
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2544..2847
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2848..3051
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3002..3022
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3096..3115
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3096..3114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3142..3151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3148..3213
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             3238..3264
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5503..5624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5540..5559)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5594..5613
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5631..6086)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5768..5789)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5980..5999
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6113..6217
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6219..6232,1..344)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(6219..6232,1..178)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
ORIGIN
        1 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
       61 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
      121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
      181 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
      241 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
      301 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
      361 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      421 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
      481 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      541 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
      601 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
      661 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
      721 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
      781 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
      841 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
      901 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
      961 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1021 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1081 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1141 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     1201 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1261 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1321 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     1381 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     1441 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     1501 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     1561 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     1621 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1681 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1741 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1801 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1861 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1921 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1981 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2041 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2101 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2161 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2221 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2281 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2341 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2401 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2461 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     2521 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2581 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2641 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2701 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2761 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2821 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2881 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2941 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3001 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     3061 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
     3121 caagctctag cgctaccggt cgccaccatg gacgcacaaa cacgacgacg tgagcgtcgc
     3181 gctgagaaac aagctcaatg gaaagctgca aacggtggat ctcctccaca tatggcttac
     3241 ccatacgatg ttccagatta cgctcctcca tctcgagctc aagcttccat gatgttccga
     3301 gaccaggtgg gcatcctcgc tggctggttc aaaggctgga atgagtgtga gcagacagtg
     3361 gccctcctgt cacttctgaa acgggtcacc cgtacccagg cccgcttcct gcagctctgc
     3421 ctggagcact cactggcgga ctgcaatgac atccacctgc tggagtcgga ggccaacagt
     3481 gctgccatcg tcagccagtg gcagcaggag tccaaagaga aggtggtgtc cctcctgctg
     3541 tcccaccttc ccctgcttca gccaggcaac acagaggcca agtcggagta catgaggcta
     3601 ctgcagaaag tgctggccta ctcaatcgag agcaatgctt tcatcgagga gagtcgccag
     3661 ctgctttcct atgccctcat ccacccagcc accacactgg aggaccgcaa cgcactggcc
     3721 ctctggctga gccacctgga agagcggttg gctagtggct tccgctcccg gccagagccc
     3781 tcctaccatt cacgtcaagg ctcagatgag tgggggggcc ctgcagagct aggccctggg
     3841 gaggcagggc caggctggca ggacaagcca ccccgggaaa atggacacgt gcccttccac
     3901 ccatccagct cagtgccgcc agccatcaac agtattggga gcaatgcaaa cacaggtctc
     3961 ccctgccaaa tccaccctag cccactgaag cgctccatgt cactcatccc tacaagcccc
     4021 caggtccctg gtgagtggcc gagtccagag gagcttgggg cccgggctgc ttttaccacg
     4081 cccgatcacg cacctctctc gccccagagc agcgtggcct cctctggcag tgagcagaca
     4141 gaggagcagg gctccagccg gaacaccttc caggaggatg gcagtggcat gaaagatgtg
     4201 ccctcatggc tcaagagcct ccgtttgcac aagtatgcag ccctcttctc acagatgagc
     4261 tacgaggaga tgatgacact gactgagcag cacctggagt ctcagaacgt caccaaaggt
     4321 gcccgccaca agatagccct gagcatccag aagctgcgtg agagacagag cgtcctcaag
     4381 tccctagaga aggatgtgct ggaaggcggg aacctacgaa acgctctgca ggagctgcag
     4441 cagatcatca tcactcccat caaggcctac agtgtcctcc aggccaccgt ggctgccgcc
     4501 accaccaccc ctactgccaa ggatggggcc ccgggggaac caccgctgcc aggtgctgag
     4561 cctcccctag cccaccccgg cacagacaaa ggcaccgagg ccaaggaccc tccagctgtg
     4621 gagaactacc cacctccacc agctccagct cccactgatg gcagtgagcc tgccccggct
     4681 cccgtcgccg acggagacat ccccagccag tttacacggg tgatgggcaa agtgtgcacc
     4741 caactgctgg tgtcccgacc agacgaggag aacatcacca gttacctcca gctcatcgaa
     4801 aagtgcctga ctcatgaggc tttcacggag acgcagaaga aacggctgct atcctggaaa
     4861 cagcaagtgc tgaagctcct ccggacattc ccgcgcaaag ccgcactaga gatgcagaac
     4921 taccggcagc agaaaggctg ggcattcggc tccaactcgc tccccatagc tggctctgtg
     4981 gggatgggag tggcccggcg tacccagcgg cagttcccaa tgcctccccg ggccctccca
     5041 cccggccgga tgggcctcct gagcccctcg ggcattgggg gtgtctcccc tcgacatgcc
     5101 ctcaccagcc ccagccttgg aggccagggc cgacagaacc tgtggtttgc caaccctgga
     5161 ggcagcaaca gcatgcccag tcagagccgc agctctgtgc agcgcaccca ctcgctcccg
     5221 gtccactcgt caccccaggc cattctcatg ttccctccag actgcccggt ccctgggcct
     5281 gacctggaga tcaatcccac tctggagtct ctgtgtctga gcatgacaga acacgccttg
     5341 ggtgatggga cagacaaaac ctccaccatc tgaggatcca ccggatctag ataactgatc
     5401 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     5461 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     5521 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5581 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     5641 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     5701 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     5761 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5821 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5881 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5941 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     6001 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     6061 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     6121 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6181 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa ga
//
