LOCUS       Exported                7149 bp ds-DNA     circular SYN 17-JUL-2012
DEFINITION  plasmid HIV Alex George Enhanced.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE-CMV-eGFP-W
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7149)
  AUTHORS   Self
  JOURNAL   Unpublished.
REFERENCE   2  (bases 1 to 7149)
  AUTHORS   UNKNOWN
  TITLE     Direct Submission
  JOURNAL   Exported Dec 19, 2022 from SnapGene 2.7.3
            http://www.snapgene.com
COMMENT     HRST Ligated into PUC248|WPRE fixed248|Changed cPPT to include SpeI 
            site for cloning promoters248|cloned in clean CMV promoter without 
            extraneous sequence248|9/19/03 Completely sequenced with 4x coverage
            to confirm perfect sequence248|11/13/03 Confirmed shortened cloning 
            to add SmaI/PacI and tag248|11/20/03 Tested Virus production and 
            expression from viral integration in parallell with all Intermediate
            Vectors and found good titer and expression
FEATURES             Location/Qualifiers
     source          1..7149
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..636
                     /gene="LTR"
                     /note="LTR"
                     /note="Long Terminal Repeat"
     misc_feature    1..453
                     /note="HIV U3"
                     /note="Full Length HIV U3"
     primer_bind     complement(158..179)
                     /note="pHAGE-A0050-8050"
     primer_bind     401..421
                     /note="pHAGE-S501-1000"
     misc_feature    454..551
                     /note="HIV R"
                     /note="HIV Repeat Region"
     misc_feature    552..636
                     /note="HIV U5"
     misc_feature    636..658
                     /note="PBS-K"
                     /note="tRNA binding site for Lysine tRNA"
     misc_feature    637..1155
                     /gene="psi"
                     /note="PSI"
                     /note="PSI Packaging Sequence"
     primer_bind     complement(667..690)
                     /note="pHAGE-A0550-0050"
     misc_feature    697..806
                     /note="HIV-PSI"
     misc_feature    807..1144
                     /note="HR-ePSI"
     primer_bind     898..919
                     /note="pHAGE-S1001-1500"
     primer_bind     complement(1166..1189)
                     /note="pHAGE-A1050-0550"
     misc_feature    1303..1536
                     /gene="rre"
                     /note="RRE"
                     /note="Rev Responsive Element"
     primer_bind     1391..1417
                     /note="pHAGE-S1501-2000"
     primer_bind     complement(1650..1678)
                     /note="pHAGE-A1550-1050"
     primer_bind     1898..1922
                     /note="pHAGE-S2001-2500"
     misc_feature    2034..2211
                     /gene="cppt"
                     /note="HIV cpPu (Trip)"
                     /note="Central Poly Purine Track"
     primer_bind     complement(2084..2112)
                     /note="pHAGE-A2050-1550"
     misc_feature    2218..2802
                     /gene="CMVp"
                     /note="CMVp"
                     /note="Cytomegalovirus Promoter"
     primer_bind     2393..2417
                     /note="pHAGE-S2501-3000"
     primer_bind     complement(2632..2655)
                     /note="pHAGE-A2550-2050"
     TATA_signal     2722..2726
                     /note="CMV Promoter TATA Box"
     primer_bind     2746..2766
                     /note="pHAGE 5' CDS"
     prim_transcript 2754
                     /note="Transcriptional Start"
                     /note="CMV Transcriptional Start Site"
     CDS             2812..3531
                     /gene="eGFP"
                     /note="eGFP"
                     /note="Enhanced Green Fluorescent Protein (Mammalian Codon 
                     Optimized)"
     primer_bind     2901..2918
                     /note="pHAGE-S3001-3500"
     primer_bind     complement(3144..3164)
                     /note="pHAGE-A3050-2550"
     primer_bind     3399..3420
                     /note="pHAGE-S3501-4000"
     misc_feature    3538..4129
                     /gene="WPRE"
                     /note="WPRE"
                     /note="Woodchuck Hepatitis Virus Posttranscriptional 
                     Regulatory Element"
     primer_bind     complement(3566..3600)
                     /note="pHAGE 3' CDS"
     primer_bind     complement(3644..3667)
                     /note="pHAGE-A3550-3050"
     primer_bind     3797..3817
                     /note="pHAGE-S4001-4500"
     primer_bind     4090..4113
                     /note="oligo #91 sequencing sense"
     primer_bind     complement(4125..4147)
                     /note="pHAGE-A4050-3550"
     misc_feature    4135..4223
                     /note="HIV nef"
     misc_feature    4185..4199
                     /note="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    4200..4435
                     /gene="dU3 LTR"
                     /note="dU3 LTR"
                     /note="Deleted U3 Long-Terminal Repeat"
     misc_feature    4237..4352
                     /note="HIV R"
                     /note="HIV Repeat Region"
     modified_base   4248
                     /note="Difference from 5' LTR"
                     /note="This T is a C in the 5' LTR"
     misc_feature    4353..4435
                     /note="HIV U5"
     primer_bind     4433..4464
                     /note="pHAGE-S4501-5000"
     misc_feature    4436..4506
                     /note="3' Flank"
                     /note="Chromosomal 3' Flanking Sequence carried over from 
                     original HIV integration site"
     primer_bind     4539..4561
                     /note="pHAGE-S6001-6500"
     promoter        4710..4744
                     /note="P-Bla"
                     /note="Beta Lactamase Promoter"
     CDS             4763..5602
                     /gene="AMPr"
                     /note="AMPr"
                     /note="Ampicillin Resistance Gene"
     primer_bind     complement(4812..4835)
                     /note="pHAGE-A6050-5550"
     primer_bind     5012..5036
                     /note="pHAGE-S6501-7000"
     primer_bind     complement(5281..5304)
                     /note="pHAGE-A6550-6050"
     primer_bind     5532..5556
                     /note="pHAGE-S7001-7500"
     primer_bind     complement(5792..5813)
                     /note="pHAGE-A7050-6550"
     rep_origin      5831..6364
                     /gene="ORI"
                     /note="ORI"
                     /note="Bacterial Origin of Replication"
     primer_bind     6036..6060
                     /note="pHAGE-S7501-8000"
     primer_bind     complement(6303..6324)
                     /note="pHAGE-A7550-7050"
     primer_bind     6539..6558
                     /note="pHAGE-S8001-8500"
     misc_feature    6601..6724
                     /gene="SV40 ori"
                     /note="SV40 ori"
     primer_bind     complement(6779..6805)
                     /note="pHAGE-A8050-7550"
     primer_bind     7044..7072
                     /note="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 gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2761 gagacgccat ccacgctgtt ttgacctcca tagaagacac cggcggccgc catggtgagc
     2821 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2881 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     2941 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3001 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3061 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3121 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3181 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3241 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     3301 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     3361 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3421 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3481 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aggatcctaa
     3541 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3601 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3661 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3721 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3781 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3841 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3901 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     3961 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4021 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4081 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg agatccttta
     4141 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     4201 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     4261 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     4321 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     4381 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     4441 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     4501 gagaggcccg ggttaattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt
     4561 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4621 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4681 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4741 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4801 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4861 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4921 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4981 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5041 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5101 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5161 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5221 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5281 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5341 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5401 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5461 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5521 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5581 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5641 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5701 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5761 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5821 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5881 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5941 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6001 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6061 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6421 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6481 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6541 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     6601 caagctcatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     6661 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     6721 cccgtggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6781 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     6841 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
     6901 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6961 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact
     7021 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg
     7081 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa
     7141 cttagtagt
//
