LOCUS       Exported                7745 bp ds-DNA     circular SYN 17-FEB-2017
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7745)
  AUTHORS   Pazour Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Dec 10, 2020 from SnapGene 5.2.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7745
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     CDS             complement(459..470)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 458"
                     /translation="IEGR"
     LTR             724..904
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     promoter        1037..1141
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             1142..2002
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 1142 .. 1210 / #ccffcc / signal sequence
                        2: 1211 .. 2002 / #ccffcc
                     Cleavage site after base 1210"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1495..1513)
                     /label=Fragment 3.REV
                     /note="color: black; sequence: GGCAGCACTGCATAATTCT"
     primer_bind     1495..1513
                     /label=Fragment 2.REV
                     /note="color: black; sequence: AGAATTATGCAGTGCTGCC"
     rep_origin      2173..2761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     promoter        3208..3238
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 3208 .. 3213 / #ffffff / -35
                        2: 3214 .. 3231 / #ffffff
                        3: 3232 .. 3238 / #ffffff / -10"
     protein_bind    3246..3262
                     /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)."
                     /note="color: #31849b"
     primer_bind     3270..3286
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     LTR             3547..4180
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    4227..4352
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    4849..5082
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
                     /note="color: #ffe4c4"
     misc_feature    5607..5724
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
     enhancer        5776..6079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        6080..6283
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             6385..6450
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /note="color: #cc99b2
                     Cleavage site after base 6450"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6426..6452)
                     /label=Fragment 2.FOR
                     /note="color: black; sequence: 
                     cGGATCttacttactAaGCGGCCGCCTTGTCATCGTCATCCTTGTAATCG"
     primer_bind     6436..6477
                     /label=Ubi_F_short
                     /note="color: black; sequence: 
                     GATGACGATGACAAGGCAGATCTCAATGGTGGTGGTGGTGGG"
     CDS             6484..6711
                     /codon_start=1
                     /label=Ubiquitin without lysines
                     /note="color: #993366"
                     /translation="MQIFVRTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF
                     AGRQLEDGRTLSDYNIQRESTLHLVLRLRGG"
     primer_bind     complement(6703..6734)
                     /label=Ubi_R_short_v2
                     /note="color: black; sequence: 
                     gcGGATCttacttactAaGCTCAACCACCTCT"
     primer_bind     6715..6733
                     /label=IRES to WPRE
                     /note="color: black; sequence: 
                     CGATTACAAGGATGACGATGACAAGGCGGCCGCtTagtaagtaaGATCCg"
     misc_feature    6734..7320
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (ECMV)"
                     /note="This feature has 3 segments:
                        1: 6734 .. 7308 / #a6acb3
                        2: 7309 .. 7311 / #a6acb3 / ATG
                        3: 7312 .. 7320 / #a6acb3"
     misc_feature    6734
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
                     /note="color: #a6acb3"
     CDS             7321..7716
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /note="color: #ccffcc"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     7374..7397
                     /label=Sequences
                     /note="color: black; sequence: tacaatcaacagcatccccatctc"
     primer_bind     7498..7508
                     /label=Ubi_F_short
                     /note="color: black; sequence: 
                     GATGACGATGACAAGGCAGATCTCAATGGTGGTGGTGGTGGG"
     primer_bind     7697..7745
                     /label=Fragment 3.FOR
                     /note="color: black; sequence: 
                     ctggttatgtgtgggagggctaaCTCGAGAGATCctaatcaacctctgg"
     primer_bind     complement(7697..7745)
                     /label=WPRE to IRES
                     /note="color: black; sequence: 
                     ccagaggttgattagGATCTCTCGAGttagccctcccacacataaccag"
     primer_bind     complement(7725..7745)
                     /label=Primer 1
                     /note="color: black; sequence: ccagaggttgattagGATCTC"
     misc_feature    join(7733..7745,1..576)
                     /label=WPRE
                     /note="woodchuck posttranscriptional regulatory element"
                     /note="color: #ffffff"
ORIGIN
        1 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
       61 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
      121 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
      181 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
      241 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
      301 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
      361 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
      421 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
      481 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
      541 agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact
      601 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
      661 attcactccc aacgaagaca agatcacctg caggacaggc gcgccctgct ttttgcttgt
      721 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      781 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      841 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
      901 agcacccggg cgattaagga aagggctaga tcattcttga agacgaaagg gcctcgtgat
      961 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     1021 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1081 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1141 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1201 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1261 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1321 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1381 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1441 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1501 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1561 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1621 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1681 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1741 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1801 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     1861 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1921 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1981 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2041 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2281 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2401 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2461 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2521 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2581 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2641 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2701 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2761 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2821 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2881 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2941 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa
     3001 gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3061 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc
     3121 gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     3181 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     3241 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt
     3301 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3361 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc
     3421 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga
     3481 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
     3541 agtagttgga agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct
     3601 accacacaca aggctacttc cctgattagc agaactacac accagggcca ggggtcagat
     3661 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag
     3721 aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg
     3781 acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg
     3841 cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac
     3901 tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc
     3961 tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag
     4021 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     4081 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     4141 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
     4201 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
     4261 gcacggcaag aggcgagggg cggcgactgc agagtacgcc aaaaattttg actagcggag
     4321 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     4381 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     4441 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     4501 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     4561 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     4621 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     4681 cacagcaagc ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     4741 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     4801 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     4861 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     4921 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     4981 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     5041 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     5101 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     5161 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     5221 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     5281 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     5341 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     5401 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     5461 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     5521 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc gccgaattct
     5581 caacaatagg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     5641 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     5701 caaattacaa aaattcaaaa ttttcgggtt tattactcgg acagcagaga tccagtttgg
     5761 actagtggag ttccgcgtta cataacttac gcaaaatggc ccgcctggct gaccgcccaa
     5821 cgacccccgc ccattgacgt caataatgac caatgttccc atagtaacgc caatagggac
     5881 tttccattga cgtcaatggg tggagtattt acgcaaaact gcccacttgg cagtacatca
     5941 agtgtatcat ttgccaacaa cgccccctat tgacgtcaat gacgcaaaat ggcccgcctg
     6001 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6061 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     6121 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     6181 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     6241 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     6301 gatcgcctgg agacgccatc cacgctgttt tgacctccat tcaagacacc ggcggcctta
     6361 attaatgatc aggatcccac catggactac aaagaccatg acggtgatta taaagatcat
     6421 gacatcgatt acaaggatga cgatgacaag gcagatctca atggtggtgg tggtgggtcg
     6481 accatgcaga tcttcgtcag aacgttaacc ggtagaacca taactctaga agttgaacca
     6541 tccgatacca tcgaaaacgt tagagctaga attcaagaca gagaaggcat tccacctgat
     6601 caacaaagat tgatctttgc cggtagacag ctcgaggacg gtagaacgct gtctgattac
     6661 aacattcaga gagagtcgac cttacatctt gtcttaagac taagaggtgg ttgagcttag
     6721 taagtaagat ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
     6781 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     6841 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     6901 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     6961 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     7021 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     7081 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     7141 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     7201 tggggcctcg gtacacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     7261 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     7321 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     7381 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     7441 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     7501 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     7561 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     7621 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     7681 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaac tcgagagatc ctaatcaacc
     7741 tctgg
//
