LOCUS       Exported                6790 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gateway-compatible destination vector for mammalian expression of 
            N-terminal HA-tag fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    pMH-HA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6790)
  AUTHORS   An L, Jiang Y, Ng HH, Man EP, Chen J, Khoo US, Gong Q, Huen MS
  TITLE     Dual-utility NLS drives RNF169-dependent DNA damage responses.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Apr 4;114(14):E2872-E2881. doi: 
            10.1073/pnas.1616602114. Epub 2017 Mar 21.
  PUBMED    28325877
REFERENCE   2  (bases 1 to 6790)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6790
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      13..441
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(100..119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     310..331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(450..470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        455..784
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      635..770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     697..716
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             810..1604
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(864..883)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1474..1493
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1778..1899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1815..1834)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1869..1888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1932..1949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2103..2691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2183..2202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2862..3722)
                     /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     3485..3504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3723..3827)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3892..4268
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4270..4473
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4424..4444
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4530..4539
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4557..4583
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4584..4610
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4629..4649
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    4650..4774
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4799..4829
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4883..5563
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(4949..4968)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5883..6188
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     6067..6086
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(6229..6353)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6381..6401
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6405..6425
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6429..6449
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6462..6491
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6527..6544)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6533..6757
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
       61 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      121 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
      181 ggttccgatt tagtgcttta cggcacctct cccccaaaaa acttgattag ggtgatggtt
      241 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
      301 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
      361 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
      421 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
      481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
      541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      601 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      661 cgcccattct ccgcccctag gctgactaat tttttttatt tatgcagagg ccgaggccgc
      721 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      781 caaaaagctc cccccccggg aggtccacaa tggttgaaca agatggattg cacgcaggtt
      841 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
      901 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
      961 ccgacctgtc cggtgccctg aatgaactcc aggacgaggc agcgcggcta tcgtggctgg
     1021 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     1081 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     1141 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     1201 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     1261 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     1321 tcgccaggct caaggcgcgt atgcccgacg gcgaggatct cgtcgtgact catggcgatg
     1381 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     1441 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     1501 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     1561 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     1621 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     1681 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     1741 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     1801 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgat
     1921 ctttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1981 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2041 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2281 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2341 tccaagctgg gctgtatgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2521 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2821 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2881 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2941 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3001 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3061 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3121 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3181 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3241 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3301 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3361 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3421 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3481 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3541 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3601 cgggcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3661 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3721 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3781 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3841 aaagtgccac ctgacgtcag atcgacggat cgggagatcg gtaccgaatt cacattgatt
     3901 attgagtagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     3961 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     4021 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     4081 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     4141 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     4201 ccagtacatg accttatggg actttcctac ttggcagtac atctacggtt agtcatcgct
     4261 attaccatag tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     4321 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4381 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4441 cgtgtacggt gggaggtcta tataagcaga gctttctggc taactagaga acccactgct
     4501 tactggcacg tggaaattaa tacgacgtgg ccaccatggc ccaggtgcag ctgcagtacc
     4561 catacgacgt gccagactac gcatacccat acgacgtgcc agactacgca ctgcaggtcg
     4621 acctcgagga aaacctgtat tttcagggca caagtttgta caaaaaagct gaacgagaaa
     4681 cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa cagactacat
     4741 aatactgtaa aacacaacat atccagtcac tatggcggcc gcattaggca ccccaggctt
     4801 tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc cgtcgagatt
     4861 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     4921 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     4981 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     5041 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     5101 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     5161 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     5221 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     5281 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     5341 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     5401 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     5461 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     5521 tgagtggcag gcggggcgta atctagagga tccggcttac taaaagccag ataacagtat
     5581 gcgtatttgc gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt
     5641 atgtcaaaaa gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct
     5701 atcagttgct caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca
     5761 gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc
     5821 tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg
     5881 aaatgcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     5941 tacagagtga tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac
     6001 gtctgctgtc agataaagtc ccccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     6061 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     6121 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     6181 gaatataaat gtcaggctcc cttatacaca gccagtctgc aggtcgacca tagtgactgg
     6241 atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata
     6301 ttgatattta tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggtctcgaga
     6361 tcaaacgggc ggccgcagat ccaaaaaaga agagaaaggt agatccaaaa aagaagagaa
     6421 aggtagatcc aaaaaagaag agaaaggtag atacggccgc agaacaaaaa ctcatctcag
     6481 aagaggatct gaatggggcc gcatagtcta gaagctcgct gatcagcctc gactgtgcct
     6541 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     6601 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     6661 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     6721 aatagcaggc atgctgggga tggcccgggc tctatggctt ctgaggcgga aagaaccagc
     6781 tggggctcta
//
