LOCUS       sequence-423103-        6527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for transient expression of MSRA gene with C-terminal HA tag,
            gene flanked by att sites for gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6527)
  TITLE     Chromosome 8p-engineering reveals increased metastasis targetable by
            patient-specific synthetic lethality
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6527)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pDEST26-MSRA-C-HA
FEATURES             Location/Qualifiers
     source          1..6527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     576..600
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    697..721
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1427..1451)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1456..1482
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1513..1532)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1514..1532)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1543..1560)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1543..1559)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1552..1574)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    1964..2098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2001..2020)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2055..2074
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2224..2679
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2311..2330)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2521..2542
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2796..3153
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2796..2987
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2819..2839)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3004..3139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3204..3998
                     /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(3258..3277)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3868..3887
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4062..4110
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4151..4170)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4270..4292
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4416..4520
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4521..4589,4590..5381)
                     /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(4739..4758)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5552..6140
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6041..6060
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6294..6311
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(6385..6489)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccattattat catgacatta acctataaaa ataggcgtat tacgaggccc tttcacgcat
       61 tagatgcatg tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
      601 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
      661 ctccggactc tagcctaggc cgcggagctc gagtcaacaa gtttgtacaa aaaagttggc
      721 atgctctcgg ccacccggag ggcttgccag ctcctcctcc tccacagcct ctttcccgtc
      781 ccgaggatgg gcaactcggc ctcgaacatc gtcagccccc aggaggcctt gccgggccgg
      841 aaggaacaga cccctgtagc ggccaaacat catgtcaatg gcaacagaac agtcgaacct
      901 ttcccagagg gaacacagat ggctgtattt ggaatgggat gtttctgggg agctgaaagg
      961 aaattctggg tcttgaaagg agtgtattca actcaagttg gttttgcagg aggctatact
     1021 tcaaatccta cttataaaga agtctgctca gaaaaaactg gccatgcaga agtcgtccga
     1081 gtggtgtacc agccagaaca catgagtttt gaggaactgc tcaaggtctt ctgggagaat
     1141 cacgacccga cccaaggtat gcgccagggg aacgaccatg gcactcagta ccgctcggcc
     1201 atctacccga cctctgccaa gcaaatggag gcagccctga gctccaaaga gaactaccaa
     1261 aaggttcttt cagagcacgg cttcggcccc atcactaccg acatccggga gggacagact
     1321 ttctactatg cggaagacta ccaccagcag tacctgagca agaaccccaa tggctactgc
     1381 ggccttgggg gcaccggcgt gtcctgccca gtgggtatta aaaaataccc aactttcttg
     1441 tacaaagtgg ttgattaccc atacgatgtt ccagattacg cttgacgcgt gcatgcgacg
     1501 tcatagctct ctccctatag tgagtcgtat tataagctag gcactggccg tcgttttaca
     1561 acgtcgtgac tgggaaaact gctagcttgg gatctttgtg aaggaacctt acttctgtgg
     1621 tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt
     1681 tttaagtgta taatgtgtta aactagctgc atatgcttgc tgcttgagag ttttgcttac
     1741 tgagtatgat ttatgaaaat attatacaca ggagctagtg attctaattg tttgtgtatt
     1801 ttagattcac agtcccaagg ctcatttcag gcccctcagt cctcacagtc tgttcatgat
     1861 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     1921 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     1981 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2041 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcga
     2101 tcctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt ggctggcgta
     2161 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2221 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2281 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2341 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2401 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2461 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2521 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2581 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2641 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     2701 tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa
     2761 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     2821 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2881 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2941 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3001 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3061 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3121 ggaggctttt ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc
     3181 tcgaacttaa ggctagagcc accatgattg aacaagatgg attgcacgca ggttctccgg
     3241 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3301 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3361 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3421 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gattggctgc
     3481 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3541 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3601 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3661 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3721 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3781 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3841 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3901 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3961 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4021 gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt
     4081 cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc cgcgtatggt
     4141 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4201 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4261 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4321 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4381 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4441 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4501 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4561 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4621 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4681 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4741 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4801 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4861 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4921 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4981 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5041 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5101 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5161 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5221 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5281 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5341 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5401 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5461 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5521 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5581 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5641 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5701 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5761 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5821 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5881 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5941 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6001 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6061 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6121 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6181 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6241 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6301 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6361 cgattcatta atgcagagct tgcaattcgc gctttttcaa tattattgaa gcatttatca
     6421 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6481 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaa
//
