LOCUS       Exported                8131 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  pMVP destination vector, empty Lentivirus vector backbone w/ Blast 
            selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    IY601: pMVP/Lenti/Blast-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8131)
  AUTHORS   Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, 
            Newgard CB
  TITLE     Creation of versatile cloning platforms for transgene expression and
            dCas9-based epigenome editing.
  JOURNAL   Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 
            10.1093/nar/gky1286.
  PUBMED    30590691
REFERENCE   2  (bases 1 to 8131)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8131
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    1981..2105
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2130..2160
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2214..2873
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2280..2299)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3215..3520
                     /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     3399..3418
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3561..3685)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3737..4053
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3904..4039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4101..4148
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4167..4565
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4652..4885
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4957..5078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4994..5013)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5048..5067
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5118..5253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5273..5292)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5274..5292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5302..5319)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5302..5318)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5311..5333)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5460..5915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5547..5566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5757..5778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5941..6045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6046..6906
                     /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(6264..6283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7077..7665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7566..7585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7819..7836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7953..7974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7989..8019
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8027..8043
                     /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)."
     primer_bind     8032..8054
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8051..8067
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8051..8067
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8086..8106
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8088..8106
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgataag cttgtctaga ggatccacta gtccagtgtg gtggaattct gcagatatca
     1981 acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta
     2041 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
     2101 ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg
     2161 tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga
     2221 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     2281 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     2341 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     2401 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
     2461 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     2521 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     2581 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     2641 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     2701 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     2761 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     2821 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taaagatctg
     2881 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     2941 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     3001 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     3061 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     3121 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     3181 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     3241 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     3301 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     3361 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     3421 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     3481 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     3541 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
     3601 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     3661 gttcagcttt cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag
     3721 aggtggcgat ggttccggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3781 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3841 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3901 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3961 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4021 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4081 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     4141 aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa gaatccaccc
     4201 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     4261 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     4321 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     4381 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     4441 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     4501 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     4561 gctaagcaca attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat
     4621 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     4681 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     4741 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     4801 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     4861 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     4921 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     4981 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5041 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     5101 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5161 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     5221 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     5281 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     5341 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     5401 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     5461 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5521 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5581 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5641 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5701 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5761 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5821 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5881 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     5941 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6001 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6061 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6121 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6181 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6241 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6301 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6361 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6421 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6481 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6541 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6601 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6661 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6721 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6781 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6841 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6901 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6961 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7021 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7081 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7141 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7201 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7261 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7321 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7381 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7441 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7501 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7561 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7621 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7681 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7741 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7801 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7861 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     7921 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7981 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8041 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     8101 taaagggaac aaaagctgga gctgcaagct t
//
