LOCUS       Exported                8646 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Allows Gateway cloning of gene of interest into retroviral pMSCV 
            vector (with Hygromycin resistance).
ACCESSION   .
VERSION     .
KEYWORDS    pMSCVhyg-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8646)
  TITLE     Gateway-compatible version of retroviral pMSCVhyg vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8646)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8646
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    64..405
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             472..888
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     818..840
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     856..872
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    914..1038
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1063..1093
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1147..1806
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1213..1232)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2148..2453
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2332..2351
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2494..2618)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2671..2694)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3056..3075
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3141..4166
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             4524..5038
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5200..5216
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5224..5254)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5269..5290
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5407..5424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5578..6166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5658..5677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6337..7197)
                     /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     6960..6979
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7198..7302)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7370..7388
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7426..7448)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7548..7567
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7761..7783
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7911..7930
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             8130..8646
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc cccgccggga
       61 ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc
      121 atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc
      181 cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac
      241 tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg
      301 tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
      361 tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc
      421 gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact
      481 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
      541 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
      601 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
      661 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
      721 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
      781 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
      841 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg aattagatct
      901 ctcgaggtta tcaacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
      961 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
     1021 acatatccag tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc
     1081 ggctcgtata atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa
     1141 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1201 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1261 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1321 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     1381 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1441 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1501 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1561 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     1621 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1681 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     1741 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     1801 gcgtaaagat ctggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct
     1861 gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt
     1921 atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg
     1981 catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg
     2041 tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt
     2101 tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg
     2161 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
     2221 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata
     2281 aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
     2341 ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc
     2401 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag
     2461 gctcccttat acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta
     2521 cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca
     2581 ttttacgttt ctcgttcagc tttcttgtac aaagtggttg ataacgaatt ctaccgggta
     2641 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     2701 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     2761 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     2821 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     2881 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     2941 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     3001 tccgggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc
     3061 tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt
     3121 tcgacctgca tcccgccacc atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     3181 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     3241 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     3301 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
     3361 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
     3421 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     3481 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
     3541 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     3601 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     3661 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     3721 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     3781 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     3841 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     3901 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
     3961 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     4021 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     4081 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     4141 gcactcgtcc gagggcaaag gaatagagta gatgccgacc gaacaagagc tgatttcgag
     4201 aacgcctcag ccagcaactc gcgcgagcct agcaaggcaa atgcgagaga acggccttac
     4261 gcttggtggc acagttctcg tccacagttc gctaagctcg ctcggctggg tcgcgggagg
     4321 gccggtcgca gtgattcagg cccttctgga ttgtgttggt ccccagggca cgattgtcat
     4381 gcccacgcac tcgggtgatc tgactgatcc cgcagattgg agatcgccgc ccgtgcctgc
     4441 cgattgggtg cagatccgtc gacctgcagc caagcttatc gataaaataa aagattttat
     4501 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     4561 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
     4621 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
     4681 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
     4741 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     4801 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     4861 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
     4921 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     4981 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg
     5041 gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa
     5101 tcctgactca attagccact gttttgaatc cacatactcc aatactcctg aaatagttca
     5161 ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc
     5221 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5281 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5341 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5401 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5461 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5521 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5581 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5641 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5701 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5761 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5821 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5881 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5941 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6001 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6061 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6121 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6181 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6241 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6301 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6361 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6421 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6481 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6541 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6601 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6661 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6721 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6781 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6841 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6901 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6961 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7021 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7081 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7141 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7201 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7261 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7321 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     7381 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     7441 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     7501 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     7561 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7621 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     7681 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     7741 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt
     7801 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     7861 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     7921 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     7981 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
     8041 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     8101 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     8161 ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt
     8221 tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt
     8281 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc
     8341 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     8401 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     8461 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga
     8521 ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt
     8581 ggactcgctg atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct
     8641 ttcatt
//
