LOCUS       Exported                7806 bp ds-DNA     circular SYN 21-NOV-2018
DEFINITION  plasmid HIV Alex George Enhanced.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7806)
  AUTHORS   David Roberts
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Apr 22, 2019 from SnapGene Viewer 4.3.6
            https://www.snapgene.com
COMMENT     10/18/2018: David Roberts cloned pHAGE-EF1aL-nlsGFP-W as follows| 
            pHAGE-CMV-nlsGFP-W received from Balazs Lab as bacterial stab| 
            Miniprep DNA extraction was cut using SpeI and NotI to seperate the 
            backbone with the nlsGFP from the CMV promoter. Simultaneously, 
            pHAGE-Ef1aL-eGFP-W was cut using SpeI and NotI, seperating the Ef1aL
            promoter from the backbone. The nlsGFP backbone and the Ef1aL 
            promoter were gel extracted and ligated together at 1:2 
            (backbone:insert). Clone 1 was sent for sequencing, as shown on the 
            map. both 5' and 3' junctions were sequenced and confirmed the 
            appropriate insertion of Ef1aL into the nlsGFP backbone.|
FEATURES             Location/Qualifiers
     source          1..7806
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..636
                     /gene="LTR"
                     /label=LTR
                     /note="Long Terminal Repeat"
     misc_feature    1..453
                     /label=HIV U3
                     /note="Full Length HIV U3"
     primer_bind     complement(158..179)
                     /label=pHAGE-A0050-8050
     primer_bind     401..421
                     /label=pHAGE-S501-1000
     misc_feature    454..551
                     /label=HIV R
                     /note="HIV Repeat Region"
     misc_feature    552..636
                     /label=HIV U5
                     /note="HIV U5"
     misc_feature    636..658
                     /label=PBS-K
                     /note="tRNA binding site for Lysine tRNA"
     misc_feature    637..1155
                     /gene="psi"
                     /label=PSI
                     /note="PSI Packaging Sequence"
     primer_bind     complement(667..690)
                     /label=pHAGE-A0550-0050
     misc_feature    697..806
                     /label=HIV-PSI
     misc_feature    807..1144
                     /label=HR-ePSI
     primer_bind     898..919
                     /label=pHAGE-S1001-1500
     primer_bind     complement(1166..1189)
                     /label=pHAGE-A1050-0550
     misc_feature    1303..1536
                     /gene="rre"
                     /label=RRE
                     /note="Rev Responsive Element"
     primer_bind     1391..1417
                     /label=pHAGE-S1501-2000
     primer_bind     complement(1650..1678)
                     /label=pHAGE-A1550-1050
     primer_bind     1898..1922
                     /label=pHAGE-S2001-2500
     primer_bind     1898..1922
                     /label=Primer #67
     misc_feature    2034..2211
                     /gene="cppt"
                     /label=HIV cpPu (Trip)
                     /note="Central Poly Purine Track"
     primer_bind     complement(2084..2112)
                     /label=pHAGE-A2050-1550
     misc_feature    2218..3406
                     /label=EF1a
                     /note="Full EF1a Promoter with intron"
     primer_bind     2339..2360
                     /label=Ef1aL sequencing primer
     primer_bind     complement(2377..2398)
                     /label=Primer #434
     TATA_signal     2391..2397
     misc_feature    2421
                     /label=Transcription Start Site
     intron          2454..3396
                     /label=EF1a Intron 1
     primer_bind     3344..3369
                     /label=Primer #156
     CDS             3416..4178
                     /gene="eGFP"
                     /label=nlseGFP
                     /note="nls Fused Enhanced Green Fluorescent Protein 
                     (Mammalian Codon Optimized) with six His Tag"
     misc_feature    3416..3442
                     /label=NLS
                     /note="Nuclear Localization Signal"
     primer_bind     3529..3546
                     /label=pHAGE-S3001-3500
     primer_bind     complement(3772..3792)
                     /label=pHAGE-A3050-2550
     primer_bind     complement(3772..3792)
                     /label=Primer #81
     primer_bind     4027..4048
                     /label=pHAGE-S3501-4000
     misc_feature    4157..4174
                     /label=Six-His Tag
                     /note="Histidine Tag"
     misc_feature    4195..4786
                     /gene="WPRE"
                     /label=WPRE
                     /note="Woodchuck Hepatitis Virus Posttranscriptional 
                     Regulatory Element"
     primer_bind     complement(4223..4257)
                     /label=pHAGE 3' CDS
     primer_bind     complement(4301..4324)
                     /label=pHAGE-A3550-3050
     primer_bind     4454..4474
                     /label=pHAGE-S4001-4500
     primer_bind     complement(4782..4804)
                     /label=pHAGE-A4050-3550
     misc_feature    4792..4880
                     /label=HIV nef
                     /note="HIV nef"
     misc_feature    4842..4856
                     /label=HIV PPT
                     /note="Polypurene Tract (Not degraded by RNAseH so it can 
                     serve as a primer for + strand DNA synthesis during viral 
                     life cycle)"
     misc_feature    4857..5092
                     /gene="dU3 LTR"
                     /label=dU3 LTR
                     /note="Deleted U3 Long-Terminal Repeat"
     misc_feature    4894..5009
                     /label=HIV R
                     /note="HIV Repeat Region"
     modified_base   4905
                     /label=Difference from 5' LTR
                     /note="This T is a C in the 5' LTR"
     misc_feature    5010..5092
                     /label=HIV U5
                     /note="HIV U5"
     primer_bind     5090..5121
                     /label=pHAGE-S4501-5000
     misc_feature    5093..5163
                     /label=3' Flank
                     /note="Chromosomal 3' Flanking Sequence carried over from 
                     original HIV integration site"
     primer_bind     5196..5218
                     /label=pHAGE-S6001-6500
     promoter        5367..5401
                     /label=P-Bla
                     /note="Beta Lactamase Promoter"
     CDS             5420..6259
                     /gene="AMPr"
                     /label=AMPr
                     /note="Ampicillin Resistance Gene"
     primer_bind     complement(5469..5492)
                     /label=pHAGE-A6050-5550
     primer_bind     5669..5693
                     /label=pHAGE-S6501-7000
     primer_bind     complement(5938..5961)
                     /label=pHAGE-A6550-6050
     primer_bind     6189..6213
                     /label=pHAGE-S7001-7500
     primer_bind     complement(6449..6470)
                     /label=pHAGE-A7050-6550
     rep_origin      6488..7021
                     /gene="ORI"
                     /label=ORI
                     /note="Bacterial Origin of Replication"
     primer_bind     6693..6717
                     /label=pHAGE-S7501-8000
     primer_bind     complement(6960..6981)
                     /label=pHAGE-A7550-7050
     primer_bind     7196..7215
                     /label=pHAGE-S8001-8500
     misc_feature    7258..7381
                     /gene="SV40 ori"
                     /label=SV40 ori
     primer_bind     complement(7436..7462)
                     /label=pHAGE-A8050-7550
     primer_bind     7701..7729
                     /label=pHAGE-S1-500
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac gcccctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     2581 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag
     2641 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     2701 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     2761 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt
     2821 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     2881 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
     2941 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     3001 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa
     3061 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     3121 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     3181 ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     3241 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     3301 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     3361 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaagcgg ccgccatggc
     3421 ccccaagaag aagcgcaagg tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3481 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3541 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     3601 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     3661 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3721 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     3781 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     3841 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3901 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3961 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4021 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4081 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4141 ggacgagctg tacaagcatc atcaccatca ccattgactc gagggcccag atcctaatca
     4201 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4261 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4321 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4381 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4441 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4501 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4561 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     4621 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4681 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4741 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctgaga tcctttaaga
     4801 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     4861 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     4921 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     4981 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5041 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     5101 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     5161 aggcccgggt taattaagga aagggctaga tcattcttga agacgaaagg gcctcgtgat
     5221 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5281 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5341 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5401 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5461 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5521 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5581 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5641 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5701 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5761 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5821 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5881 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5941 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6001 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6061 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6121 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6181 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6241 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6301 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6361 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6421 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6481 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6541 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6601 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6661 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6721 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6781 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6841 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6901 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6961 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7021 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7081 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     7141 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     7201 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa
     7261 gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     7321 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc
     7381 gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     7441 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     7501 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt
     7561 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     7621 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc
     7681 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga
     7741 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
     7801 agtagt
//
