LOCUS       pSMAL                   9950 bp    DNA     circular UNA 02-OCT-2020
DEFINITION  Gateway destination vector with bidirectional CMV-SFFV promoter for
            high expression of gene of interest and GFP.
ACCESSION   urn.local...e-b8v8yua
VERSION     urn.local...e-b8v8yua
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    91..168
                     /created_by="User"
                     /label="SV40 ori"
                     /note="Geneious type: origin of replication"
     misc_feature    465..771
                     /created_by="User"
                     /modified_by="User"
                     /label="F1 phage ori"
                     /note="Geneious type: origin of replication"
     promoter        967..995
                     /created_by="User"
                     /label="AmpR promoter"
     resistance      1037..1897
                     /created_by="User"
                     /label="AmpR"
     ColE1_origin    1992..2674
                     /created_by="User"
                     /label="ColE1 origin"
     promoter        2980..3009
                     /created_by="User"
                     /label="lac promoter"
     promoter        3208..3775
                     /created_by="User"
                     /label="CMV immediate early promoter"
     LTR             3793..3976
                     /created_by="User"
                     /label="HIV-1 5' LTR"
     other           4638..4871
                     /created_by="User"
                     /label="RRE"
     misc_feature    5370..5385
                     /created_by="User"
                     /modified_by="User"
                     /label="cPPT"
                     /note="Geneious type: Central polypurine tract"
     polyA_site      complement(5508..5750)
                     /created_by="User"
                     /Transferred_From="07 TOPO revcas eGFP KpnI AgeI"
                     /Transferred_Similarity="100.00%"
                     /modified_by="vangalen"
                     /label="polyA"
     SV40_polyA      complement(5618..5749)
                     /created_by="User"
                     /modified_by="User"
                     /label="SV40 polyA"
     CDS             complement(5792..6511)
                     /created_by="User"
                     /modified_by="User"
                     /Transferred_From="07 TOPO revcas eGFP KpnI AgeI"
                     /Transferred_Similarity="100.00%"
                     /label="eGFP"
     promoter        complement(6594..6653)
                     /created_by="User"
                     /modified_by="User"
                     /Transferred_From="07 TOPO revcas eGFP KpnI AgeI"
                     /Transferred_Similarity="100.00%"
                     /label="minhCMV"
     promoter        6660..7074
                     /vntifkey=19
                     /modified_by="vangalen"
                     /label="SFFVp"
     loxP            7087..7120
                     /created_by="User"
                     /label="loxP"
     primer_bind     7104..7130
                     /%GC="37.0"
                     /Hairpin_Tm="37.8"
                     /Self_Dimer_Tm="None"
                     /Tm="60.4"
                     /annotation_group="2196504327973267c1dc43-28af-448d-8f07-a
                     91adcc23aa0"
                     /created_by="primer3"
                     /Sequence="ACATTATACGAAGTTATCCCAGCTTGA"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pSMAL"
                     /label="PvG114-loxPattB1_F"
     primer_bind     complement(7122..7141)
                     /%GC="40.0"
                     /Hairpin_Tm="45.9"
                     /Self_Dimer_Tm="7.1"
                     /Tm="53.3"
                     /annotation_group="217620513997776fd05009-69bb-449c-8f2a-d
                     0779552b3f6"
                     /created_by="primer3"
                     /Sequence="AACTTGTGATATCAAGCTGG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pSMAL"
                     /label="PvG501-attR1_R_seq"
     attR1           7136..7260
                     /Mismatches=1
                     /%_Identity="99.2"
                     /Motif="CAACTTTGTACAAAAAAGCTGAACGAGAAACGTAAAATGATATAAATATC
                     AATATATTAAATTAGATTTTGCATAAAAAACAGACTACATAATACTGTAAAACACAAC
                     ATATCCAGTCACTATGG"
                     /annotation_group="attR1: 7,136 -> 7,260"
                     /label="attR1"
     misc_feature    7368..8027
                     /created_by="User"
                     /label="CmR"
                     /note="Geneious type: Chloramphenicol resistance"
     ccdB            8381..8674
                     /created_by="User"
                     /label="ccdB"
     attR2           complement(8714..8838)
                     /Mismatches=1
                     /%_Identity="99.2"
                     /Motif="CAACTTTGTACAAGAAAGCTGAACGAGAAACGTAAAATGATATAAATATC
                     AATATATTAAATTAGATTTTGCATAAAAAACAGACTACATAATACTGTAAAACACAAC
                     ATATCCAGTCACTATGG"
                     /annotation_group="attR2: 8,714 <- 8,838"
                     /label="attR2"
     loxP            8882..8915
                     /created_by="User"
                     /modified_by="User"
                     /label="loxP"
     WPRE            8923..9498
                     /created_by="User"
                     /label="WPRE"
     primer_bind     complement(9022..9045)
                     /%GC="50.0"
                     /Hairpin_Tm="41.1"
                     /Self_Dimer_Tm="None"
                     /Tm="63.4"
                     /annotation_group="2194892486457385c19f25-d436-4a4c-8b53-b
                     0005ddbca59"
                     /created_by="primer3"
                     /Sequence="GCCATACGGGAAGCAATAGCATGA"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="pSMAL"
                     /label="PvG049-WPRE-rev"
     misc_feature    9579..9594
                     /created_by="User"
                     /modified_by="User"
                     /label="cPPT"
                     /note="Geneious type: Central polypurine tract"
     LTR             9649..9829
                     /created_by="User"
                     /label="HIV-1 5' LTR"
ORIGIN      
        1 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
       61 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc
      121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
      181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcgtc
      241 gagacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
      301 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
      361 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
      421 tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg
      481 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      541 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      601 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
      661 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
      721 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
      781 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
      841 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
      901 tttcccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      961 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1021 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1081 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1141 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1201 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1261 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1321 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1381 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1441 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1501 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1561 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1621 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1681 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1741 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1801 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1861 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1921 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1981 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2041 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2101 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2161 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2221 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2281 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2341 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2401 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2461 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2521 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2581 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2641 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2701 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2761 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2821 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2881 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2941 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     3001 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     3061 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
     3121 ttggccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
     3181 aacattaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     3241 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     3301 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     3361 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     3421 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     3481 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     3541 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     3601 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     3661 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     3721 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     3781 tcgtttagtg aaccggggtc tctctggtta gaccagatct gagcctggga gctctctggc
     3841 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     3901 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     3961 tggaaaatct ctagcagtgg cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc
     4021 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     4081 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     4141 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     4201 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     4261 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     4321 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     4381 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     4441 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     4501 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     4561 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     4621 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     4681 atgggcgcag cctcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     4741 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     4801 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     4861 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     4921 tggaatgcta gttggagtaa taaatctctg gaacagattg gaatcacacg acctggatgg
     4981 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     5041 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     5101 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     5161 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     5221 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     5281 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     5341 acggatctcg acggtatcgg ttaactttta aaagaaaagg ggggattggg gggtacagtg
     5401 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     5461 aaattacaaa aattcaaaat tttatcgatc acgagactag cctcgagtga tcataatcag
     5521 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     5581 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     5641 ttacaaataa ggcaatagca tcacaaattt cacaaataag gcattttttt cactgcattc
     5701 tagttttggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct caaatccctc
     5761 ggaagctgcg cctgtcatcg aattcaccgg tctacttgta cagctcgtcc atgccgagag
     5821 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     5881 ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     5941 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     6001 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
     6061 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     6121 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     6181 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     6241 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     6301 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     6361 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     6421 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     6481 cggtgaacag ctcctcgccc ttgctcacca tggtggtacc gaattccgcg gaggctggat
     6541 cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga
     6601 cggttcacta aacgagctct gcttatatag gcctcccacc gtacacgcct accactagta
     6661 atgaaagacc ccacctgtag gtttggcaag ctagctgcag taacgccatt ttgcaaggca
     6721 tggaaaaata ccaaaccaag aatagagaag ttcagatcaa gggcgggtac atgaaaatag
     6781 ctaacgttgg gccaaacagg atatctgcgg tgagcagttt cggccccggc ccggggccaa
     6841 gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc aagaacagat ggtccccaga
     6901 tatggcccaa ccctcagcag tttcttaaga cccatcagat gtttccaggc tcccccaagg
     6961 acctgaaatg accctgcgcc ttatttgaat taaccaatca gcctgcttct cgcttctgtt
     7021 cgcgcgcttc tgcttcccga gctctataaa agagctcaca acccctcact cggcttaatt
     7081 aagatcataa cttcgtatag catacattat acgaagttat cccagcttga tatcacaagt
     7141 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
     7201 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
     7261 cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
     7321 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     7381 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     7441 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     7501 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     7561 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     7621 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     7681 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     7741 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     7801 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     7861 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     7921 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     7981 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg
     8041 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     8101 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     8161 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     8221 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     8281 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     8341 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     8401 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     8461 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     8521 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     8581 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     8641 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     8701 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     8761 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     8821 ctttcttgta caaagtggtg atatcgaatt cctgcagccc gggggatcca ctagttctag
     8881 aataacttcg tatagcatac attatacgaa gttattcgac aatcaacctc tggattacaa
     8941 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     9001 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     9061 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     9121 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     9181 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     9241 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     9301 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     9361 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     9421 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     9481 tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacc tttaagacca
     9541 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     9601 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     9661 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     9721 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     9781 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     9841 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     9901 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
//
