LOCUS       pHAGE-EFS-MCP-HALOnls        7441 bp    DNA     circular SYN 23-AUG-2018
DEFINITION  plasmid HIV Alex George Enhanced.
ACCESSION   Your mama'
SOURCE      Unknown.
  ORGANISM  Unknown
            Unclassified.
REFERENCE   1  (bases 1 to 8541)
  AUTHORS   Self
  JOURNAL   Unpublished.
COMMENT     HRST Ligated into PUC
            WPRE fixed
            Changed cPPT to include SpeI site for cloning promoters
            cloned in clean CMV promoter without extraneous sequence
            9/19/03 Completely sequenced with 4x coverage to confirm perfect sequence
            10/08/03 Mutations found in IRES Element Between BamHI and NdeI sites
            11/03/03 After Re-cloning the IRES from TOPO vector sequence is now confirmed to be perfect (IRES contains 7As at one point which seems to only be 6As in MCV genome)
            11/13/03 Confirmed shortened cloning to add SmaI/PacI and tag
            1/27/04 Confirmed perfect second position ZsGreen sequence with multiple oligos
            5/24/04 Sequencing confirmed perfect cPPT-CMV-DsRed Express region
COMMENT     This file is created by Vector NTI
            http://www.invitrogen.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTDATE|-12434372|
COMMENT     VNTDBDATE|-12434372|
COMMENT     LSOWNER|
COMMENT     VNTNAME|pHAGE-EFS-MCP-HALOnls|
COMMENT     VNTAUTHORNAME|Demo User|
COMMENT     Vector_NTI_Display_Data_(Do_Not_Edit!)
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COMMENT        #2=(CRSite (CStringList) "BamHI" "ggatcc" 2 0 1 2901 0 0 "") 
COMMENT        #3=(CRSite (CStringList) "EcoRI" "gaattc" 2 0 1 2783 0 0 "") 
COMMENT        #4=(CRSite (CStringList) "MluI" "acgcgt" 2 0 1 2507 0 0 "") 
COMMENT        #5=(CRSite (CStringList) "NcoI" "ccatgg" 2 0 1 2527 0 0 "") 
COMMENT        #6=(CRSite (CStringList) "NheI" "gctagc" 2 0 1 2222 0 0 "") 
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COMMENT        #9=(CRSite (CStringList) "RsrII" "cggwccg" 3 0 0 0 "") 
COMMENT        #10=(CRSite (CStringList) "SacII" "ccgcgg" 5 0 1 4335 0 0 "") 
COMMENT        #11=(CRSite (CStringList) "SbfI" "cctgcagg" 7 0 0 0 "") 
COMMENT        #12=(CRSite (CStringList) "SpeI" "actagt" 2 0 0 0 "") 
COMMENT        #13=(CRSite (CStringList) "XbaI" "tctaga" 2 0 1 3825 0 0 "") 
COMMENT        #14=(CRSite (CStringList) "XhoI" "ctcgag" 2 0 1 3795 0 0 "") 
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COMMENT                          "plasmid HIV Alex George Enhanced." 1) 
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COMMENT                          "Division: SYN" 1) 
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COMMENT                          "Modification Date in the Original DB: 23-MAY-2016" 1) 
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COMMENT                          "Original Accession Number(s): <a href=\"ncbilink:dnarna/Your\">Your</a>; <a href=\"ncbilink:dnarna/mama'\">mama'</a>" 
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FEATURES             Location/Qualifiers
     misc_feature    1..636
                     /gene="LTR"
                     /vntifkey="21"
                     /label=LTR
                     /note="Long Terminal Repeat"
     misc_feature    637..1155
                     /gene="psi"
                     /product="pbs-gag"
                     /vntifkey="21"
                     /label=PSI
                     /note="PSI Packaging Sequence"
     misc_feature    1303..1536
                     /gene="rre"
                     /product="minimal RRE"
                     /vntifkey="21"
                     /label=RRE
                     /note="Rev Responsive Element"
     misc_feature    2034..2211
                     /gene="cppt"
                     /vntifkey="21"
                     /label=HIV\cpPu\(Trip)
                     /note="Central Poly Purine Track"
     misc_feature    4492..4727
                     /gene="dU3 LTR"
                     /vntifkey="21"
                     /label=dU3\LTR
                     /note="Deleted U3 Long-Terminal Repeat"
     CDS             5037..5897
                     /gene="AMPr"
                     /product="beta-lactamase (mature form)"
                     /vntifkey="4"
                     /label=AMPr
                     /note="Ampicillin Resistance Gene"
     rep_origin      6123..6656
                     /gene="ORI"
                     /product="ColE1 origin of replication"
                     /vntifkey="33"
                     /label=ORI
                     /note="Bacterial Origin of Replication"
     misc_feature    6893..7016
                     /gene="SV40 ori"
                     /vntifkey="21"
                     /label=SV40\ori
     misc_feature    4427..4515
                     /vntifkey="21"
                     /label=HIV\nef
                     /note="HIV nef"
     misc_feature    454..551
                     /vntifkey="21"
                     /label=HIV\R
                     /note="HIV Repeat Region"
     misc_feature    4529..4644
                     /vntifkey="21"
                     /label=HIV\R
                     /note="HIV Repeat Region"
     misc_feature    4645..4727
                     /vntifkey="21"
                     /label=HIV\U5
                     /note="HIV U5"
     modified_base   4540..4540
                     /vntifkey="23"
                     /label=Difference\from\5'\LTR
                     /note="This T is a C in the 5' LTR"
     misc_feature    636..658
                     /vntifkey="21"
                     /label=PBS-K
                     /note="tRNA binding site for Lysine tRNA"
     misc_feature    697..806
                     /vntifkey="21"
                     /label=HIV-PSI
     misc_feature    807..1144
                     /vntifkey="21"
                     /label=HR-ePSI
     misc_feature    4728..4798
                     /vntifkey="21"
                     /label=3'\Flank
                     /note="Chromosomal 3' Flanking Sequence carried over from original HIV integration site"
     misc_feature    552..636
                     /vntifkey="21"
                     /label=HIV\U5
                     /note="HIV U5"
     misc_feature    1..453
                     /vntifkey="21"
                     /label=HIV\U3
                     /note="Full Length HIV U3"
     bHLH_domain     4938..5036
                     /vntifkey="1"
                     /label=Bla\promoter
     misc_feature    2028..2033
                     /vntifkey="21"
                     /label=(EcoRI/MfeI)
     misc_feature    3830..4421
                     /vntifkey="21"
                     /label=WPRE
     misc_feature    2240..2490
                     /vntifkey="21"
                     /label=EFS\promoter
     misc_feature    2492..2497
                     /vntifkey="21"
                     /label=(BspEI/AgeI)
     misc_feature    2528..2878
                     /vntifkey="21"
                     /label=MCP
     misc_feature    3800..3820
                     /vntifkey="21"
                     /label=nls
     misc_feature    2906..3793
                     /vntifkey="21"
                     /label=HaloTag
BASE COUNT     1908 a      1786 c      1933 g      1814 t 
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 ttgacaaatg 
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat 
     2221 gctagctgat gcgggccctt cgagtggctc cggtgcccgt cagtgggcag agcgcacatc 
     2281 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag 
     2341 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg 
     2401 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt 
     2461 tgccgccaga acacaggtgt cgtgacgcgg ttccggttaa ggtgaacgcg tgcaggctgg 
     2521 cgccaccatg gcttctaact ttactcagtt cgttctcgtc gacaatggcg gaactggcga 
     2581 cgtgactgtc gccccaagca acttcgctaa cgggatcgct gaatggatca gctctaactc 
     2641 gcgttcacag gcttacaaag taacctgtag cgttcgtcag agctctgcgc agaatcgcaa 
     2701 atacaccatc aaagtcgagg tgcctaaagg cgcctggcgt tcgtacttaa atatggaact 
     2761 aaccattcca attttcgcca cgaattccga ctgcgagctt attgttaagg caatgcaagg 
     2821 tctcctaaaa gatggaaacc cgattccctc agcaatcgca gcaaactccg gcatctacgg 
     2881 gggatctggt agcggcgctg gatccgcaga aatcggtact ggctttccat tcgaccccca 
     2941 ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac 
     3001 ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc 
     3061 gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga 
     3121 caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga 
     3181 agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt 
     3241 ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg 
     3301 ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg 
     3361 caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct 
     3421 gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt 
     3481 cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg 
     3541 tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc 
     3601 tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc 
     3661 tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct 
     3721 gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtcgacgct 
     3781 ggagatttcc ggcctcgagc caaagaaaaa gcggaaagtg taatctagat aatcaacctc 
     3841 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc 
     3901 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca 
     3961 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg 
     4021 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca 
     4081 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg 
     4141 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg 
     4201 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg 
     4261 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg 
     4321 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc 
     4381 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tgagatcctt taagaccaat 
     4441 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 
     4501 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt 
     4561 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 
     4621 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 
     4681 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 
     4741 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc 
     4801 cgggttaatt aaggaaaggg ctagatcatt cttgaagacg aaagggcctc gtgatacgcc 
     4861 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc 
     4921 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 
     4981 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga 
     5041 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 
     5101 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag 
     5161 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 
     5221 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg 
     5281 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 
     5341 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 
     5401 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag 
     5461 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 
     5521 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 
     5581 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc 
     5641 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 
     5701 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 
     5761 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 
     5821 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 
     5881 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa 
     5941 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 
     6001 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 
     6061 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 
     6121 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 
     6181 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 
     6241 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 
     6301 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 
     6361 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 
     6421 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 
     6481 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 
     6541 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 
     6601 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 
     6661 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 
     6721 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 
     6781 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 
     6841 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agcaagctca 
     6901 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt 
     6961 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccgtggc 
     7021 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 
     7081 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 
     7141 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttcacaa 
     7201 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 
     7261 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc 
     7321 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct 
     7381 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag 
     7441 t 
//
