LOCUS       sequence-288978-        7609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg7_1-632.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7609)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7609)
  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: pCIneo-lambdaN-HA-HsSmg7_1-632_I
FEATURES             Location/Qualifiers
     source          1..7609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1098..1107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1104..1169
                     /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             1194..1220
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3313..3434
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3324..3343)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3378..3397
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3620..4075
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3707..3726)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3917..3938
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4192..4549
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4192..4383
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4215..4235)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4400..4535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4600..5394
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4654..4673)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5264..5283
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5458..5506
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5547..5566)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5666..5688
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5726..5744)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5812..5916
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5917..5985,5986..6777)
                     /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     complement(6135..6154)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6948..7536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7437..7456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc aattgcggcc accatggacg cacaaacacg acgacgtgag cgtcgcgctg
     1141 agaaacaagc tcaatggaaa gctgcaaacg gtggatctcc tccacatatg gcttacccat
     1201 acgatgttcc agattacgct cctccactcg agatgagcct gcagagcgcg cagtacctcc
     1261 ggcaggcaga agtcctgaag gctgacatga cagattctaa gctgggtcca gctgaagtct
     1321 ggacatccag gcaggctctg caggacctgt accagaaaat gctagttacc gatttggaat
     1381 acgctttaga caagaaagta gaacaggatc tctggaatca cgcctttaag aatcagatca
     1441 caacactaca aggccaggca aagaatcgag caaatccgaa tcggagtgaa gttcaggcaa
     1501 acctttctct gttcctagag gcagctagtg gcttctatac tcagttatta caagaactgt
     1561 gtacagtatt taatgtagat ttaccatgcc gtgtgaagtc ttcccaattg ggaattatca
     1621 gcaataaaca gacgcatacc agcgccatag tgaagccaca gtctagctcc tgttcctata
     1681 tctgccagca ctgcctcgtc caccttggag acattgctcg atacagaaac cagaccagcc
     1741 aggcagagtc ctactatagg catgcagctc agcttgtccc ctccaatggt cagccttata
     1801 atcagttggc tatcttagct tcttccaaag gagaccatct gaccacaatt ttctactact
     1861 gcagaagcat tgctgtgaag ttccctttcc cagctgcctc cactaatctg caaaaagcac
     1921 tttctaaagc actggaaagc cgagatgagg tgaaaaccaa gtggggtgtt tctgacttca
     1981 tcaaggcctt tattaaattc cacggtcatg tgtacctgag taagagcttg gaaaagttga
     2041 gccctcttcg agagaaattg gaagaacagt ttaagaggct gctattccaa aaagctttca
     2101 actctcagca gttagttcat gtcactgtca ttaacctgtt tcaacttcat caccttcgtg
     2161 actttagcaa tgaaaccgag cagcacactt atagccaaga tgagcagcta tgttggacac
     2221 agttgctggc cctctttatg tcttttcttg gcatcctgtg caagtgtcct ctacagaatg
     2281 agtctcagga ggagtcctac aatgcctatc ctcttccagc agtcaaggtc tccatggact
     2341 ggctaagact cagacccagg gtctttcagg aggcagtggt ggatgaaaga cagtacattt
     2401 ggccctggtt gatttccctt ctgaatagtt tccatcccca tgaagaggac ctctcaagta
     2461 ttagtgcgac accacttcca gaggagtttg aattacaagg atttttggca ttgagacctt
     2521 ctttcaggaa cttggatttt tccaaaggtc accagggtat tacaggggac aaagaaggcc
     2581 agcaacgacg aatacgacag caacgcttga tctctatagg caaatggatt gctgataatc
     2641 agccaaggct gattcagtgt gaaaatgagg tagggaaatt gttgtttatc acagaaatcc
     2701 cagaattaat actggaagac cccagtgaag ccaaagagaa cctcattctg caggaaacat
     2761 ctgtgataga gtcgctggct gcagatggga gcccagggct aaaatcagtg ctatctacaa
     2821 gccgaaattt aagcaacaac tgtgacacag gagagaagcc agtggttacc ttcaaagaaa
     2881 acattaagac acgagaagtg aacagagacc aaggaagaag ttttcctccc aaagaggtga
     2941 gaagggacta tagcaaagga ataactgtaa ctaagaatga tggaaagaag gacaacaaca
     3001 agaggaaaac tgaaaccaag aaatgcacct tagaaaagtt acaggaaaca ggaaagcaga
     3061 atgtggcagt gcaggtaaaa tcccagacag aactaagaaa gactccagtg tctgaagcca
     3121 gaaaaacacc tgtaactcaa accccaactc aagcaagtaa ctcccagttc atccccattc
     3181 atcaccctgg agccttccct cctcttccca gcaggccagg gtttccgccc ccaacatatg
     3241 ttatcccccc gcctgtggca ttttctgcgg ccgcttccct ttagtgaggg ttaatgcttc
     3301 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     3361 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     3421 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     3481 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc cgataaggat
     3541 cgatccgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3601 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3661 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3721 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3781 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3841 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3901 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3961 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4021 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     4081 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc
     4141 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     4201 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4261 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4321 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4381 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4441 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4501 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgattc
     4561 ttctgacaca acagtctcga acttaaggct agagccacca tgattgaaca agatggattg
     4621 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4681 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4741 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4801 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4861 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4921 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4981 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5041 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5101 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5161 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5221 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5281 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5341 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5401 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga tggccgcaat
     5461 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata
     5521 aggatccgcg tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5581 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5641 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5701 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5761 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5821 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5881 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5941 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6001 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6061 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6121 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6181 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6241 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6301 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6361 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6421 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6481 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6541 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6601 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6661 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6721 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6781 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6841 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6901 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7081 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7501 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7561 cggttcctgg ccttttgctg gccttttgct cacatggctc gacagatct
//
