LOCUS       Exported                7076 bp ds-DNA     circular SYN 01-AUG-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV_GR-RE_MLP_BC1094-luc2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7076)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 17, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     LOCUS       pAAVspace_DEST_luc2_WPRE      8137 bp    DNA   CIRCULAR 
                  07-01-2013

            DEFINITION  Gateway Parent Destination Vector, AAV destination 
            vector with firefly luciferase ORF

            ACCESSION   V825

            KEYWORDS    

            SOURCE      

              ORGANISM  

            REFERENCE   1 - 8137 (bases 1 to 8137)

              AUTHORS   

              JOURNAL   

            COMMENT
FEATURES             Location/Qualifiers
     source          1..7076
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          287..1197
                     /label=seq verified pAAV
                     /note="seq verified pAAV"
     source          1185..1188
                     /label=AGCT inserted
                     /note="AGCT inserted"
     misc_feature    0
                     /label=Untitled
     misc_feature    0
                     /label=Untitled
     misc_feature    1..183
                     /label=ITR
                     /note="ITR"
     protein_bind    484..508
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     enhancer        509..580
                     /label=4x GR-RE
                     /note="4x GR-RE"
     enhancer        509..526
                     /label=GR-RE
                     /note="GR-RE"
     variation       510..524
                     /label=core consensus
                     /note="core consensus"
     enhancer        527..544
                     /label=GR-RE
                     /note="GR-RE"
     variation       528..542
                     /label=core consensus
                     /note="core consensus"
     enhancer        545..562
                     /label=GR-RE
                     /note="GR-RE"
     variation       546..560
                     /label=core consensus
                     /note="core consensus"
     enhancer        563..580
                     /label=GR-RE
                     /note="GR-RE"
     variation       564..578
                     /label=core consensus
                     /note="core consensus"
     misc_feature    581..601
                     /label=spacer 1
                     /note="spacer 1"
     enhancer        602..673
                     /label=4x GR-RE
                     /note="4x GR-RE"
     enhancer        602..619
                     /label=GR-RE
                     /note="GR-RE"
     variation       603..617
                     /label=core consensus
                     /note="core consensus"
     enhancer        620..637
                     /label=GR-RE
                     /note="GR-RE"
     variation       621..635
                     /label=core consensus
                     /note="core consensus"
     enhancer        638..655
                     /label=GR-RE
                     /note="GR-RE"
     variation       639..653
                     /label=core consensus
                     /note="core consensus"
     enhancer        656..673
                     /label=GR-RE
                     /note="GR-RE"
     variation       657..671
                     /label=core consensus
                     /note="core consensus"
     misc_feature    674..694
                     /label=spacer 2
                     /note="spacer 2"
     enhancer        695..766
                     /label=4x GR-RE
                     /note="4x GR-RE"
     enhancer        695..712
                     /label=GR-RE
                     /note="GR-RE"
     variation       696..710
                     /label=core consensus
                     /note="core consensus"
     enhancer        713..730
                     /label=GR-RE
                     /note="GR-RE"
     variation       714..728
                     /label=core consensus
                     /note="core consensus"
     enhancer        731..748
                     /label=GR-RE
                     /note="GR-RE"
     variation       732..746
                     /label=core consensus
                     /note="core consensus"
     enhancer        749..766
                     /label=GR-RE
                     /note="GR-RE"
     variation       750..764
                     /label=core consensus
                     /note="core consensus"
     protein_bind    complement(767..787)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        838..878
                     /label=Adenovirus major late promoter
                     /note="Adenovirus major late promoter"
     protein_bind    928..948
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     949..974
                     /label=Dec1 #10243
                     /note="Dec1 #10243"
     promoter        975..994
                     /label=T3
                     /note="T3"
     promoter        975..993
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        995..1007
                     /label=truncated SP6
                     /note="truncated SP6"
     misc_feature    1008..1056
                     /label=E01094
     promoter        complement(1057..1075)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1076..1101)
                     /label=Dec2 #9752
                     /note="Dec2 #9752"
     protein_bind    complement(1099..1123)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1202..1211
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1208..2860
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTITEKEIVDYVASQATTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    1318
                     /label=modified base, silent mutation, C--&gt;T
                     /note="modified base, silent mutation, C-->T"
     misc_feature    2686
                     /label=modified base
                     /note="modified base, point mutation, atg --> atA (Met --> 
                     Ile)"
     misc_feature    2724
                     /label=modified base
                     /note="modified base, point mutation, gtt --> gCt (Val --> 
                     Ala)"
     misc_feature    2882..3474
                     /label=WPRE
                     /note="WPRE"
     misc_feature    2888..3476
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3359..3370)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    3493..3761
                     /label=bGH_PA_terminator
                     /note="bGH_PA_terminator"
     polyA_signal    3509..3733
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3771..3953
                     /label=ITR
                     /note="ITR"
     promoter        complement(3954..4283)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3968..4103
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4470..5103
                     /direction=RIGHT
                     /label=pUC
                     /note="pUC"
     rep_origin      complement(4529..5117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5288..6148)
                     /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"
     promoter        complement(6149..6253)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7041..7059
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 tagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggtacc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      241 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      301 catcctgggg cacaagctgg agtacaacta caacagccac aacgtccata tcatggccga
      361 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
      421 cgtgcagctc gccgaccact catatgtact cgagttgtcg acggtatcga taagcttgat
      481 atcacaagtt tgtacaaaaa agcaggctgg gaacattatg tcctgtggga acagtatgtc
      541 ctgagggaac attatgtcct gtgggaacat tatgtcctgt acgcgtggcc tcgcctgcag
      601 ggggaacatt atgtcctgtg ggaacagtat gtcctgaggg aacattatgt cctgtgggaa
      661 cattatgtcc tgtggcctcc tgcaggaggc ggccgggaac attatgtcct gtgggaacag
      721 tatgtcctga gggaacatta tgtcctgtgg gaacattatg tcctgtcaac ttttctatac
      781 aaagttggca agtgcaggtg ccagaacatt tctctggcct aactggaaga cgctagcggg
      841 gggctataaa agggggtggg ggcgttcgtc ctcactctag atctgcgatc taagtaagct
      901 tggcaatccg gtactgttgg taaagcccaa ctttgtataa taaagttgag ctagttgcta
      961 agtctgccga gtagaattaa ccctcactaa agggtaggtg acactataat ctctacttta
     1021 catacataca ggcagtacat acattacttc tattacccta tagtgagtcg tattagtaga
     1081 cgcgagtcaa tgcatgtacg acagctttct tgtacaaagt ggtgatatcg aattcctgca
     1141 gcccggggga tccgcccggg ctagagcggc cgccaccgcg gtggagctcg cgactagtga
     1201 cgccaccatg gaagatgcca aaaacattaa gaagggccca gcgccattct acccactcga
     1261 agacgggacc gccggcgagc agctgcacaa agccatgaag cgctacgccc tggtgcctgg
     1321 caccatcgcc tttaccgacg cacatatcga ggtggacatt acctacgccg agtacttcga
     1381 gatgagcgtt cggctggcag aagctatgaa gcgctatggg ctgaatacaa accatcggat
     1441 cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat
     1501 cggtgtggct gtggccccag ctaacgacat ctacaacgag cgcgagctgc tgaacagcat
     1561 gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa
     1621 cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc atggatagca agaccgacta
     1681 ccagggcttc caaagcatgt acaccttcgt gacttcccat ttgccacccg gcttcaacga
     1741 gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga tcatgaacag
     1801 tagtggcagt accggattgc ccaagggcgt agccctaccg caccgcaccg cttgtgtccg
     1861 attcagtcat gcccgcgacc ccatcttcgg caaccagatc atccccgaca ccgctatcct
     1921 cagcgtggtg ccatttcacc acggcttcgg catgttcacc acgctgggct acttgatctg
     1981 cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc gcagcttgca
     2041 agactataag attcaatctg ccctgctggt gcccacacta tttagcttct tcgctaagag
     2101 cactctcatc gacaagtacg acctaagcaa cttgcacgag atcgccagcg gcggggcgcc
     2161 gctcagcaag gaggtaggtg aggccgtggc caaacgcttc cacctaccag gcatccgcca
     2221 gggctacggc ctgacagaaa caaccagcgc cattctgatc acccccgaag gggacgacaa
     2281 gcctggcgca gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg acttggacac
     2341 cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat
     2401 gagcggctac gttaacaacc ccgaggctac aaacgctctc atcgacaagg acggctggct
     2461 gcacagcggc gacatcgcct actgggacga ggacgagcac ttcttcatcg tggaccggct
     2521 gaagagcctg atcaaataca agggctacca ggtagcccca gccgaactgg agagcatcct
     2581 gctgcaacac cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg
     2641 cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa accataaccg agaaggagat
     2701 cgtggactat gtggccagcc aggctacaac cgccaagaag ctgcgcggtg gtgttgtgtt
     2761 cgtggacgag gtgcctaaag gactgaccgg caagttggac gcccgcaaga tccgcgagat
     2821 tctcattaag gccaagaagg gcggcaagat cgccgtgtaa ttcgaattcg atatcaagct
     2881 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2941 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3001 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3061 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3121 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3181 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3241 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3301 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3361 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3421 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3481 ataccgtcga ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3541 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3601 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3661 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3721 gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgactag agcatggcta
     3781 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     3841 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3901 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagag ctttttgcaa
     3961 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     4021 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     4081 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     4141 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     4201 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     4261 tttccacacc ctaactgaca cacattccac agctgcatta atgaatcggc caacgcgcgg
     4321 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4381 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4441 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4501 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4561 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4621 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4681 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4741 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4801 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4861 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4921 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4981 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5041 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5101 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5161 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5221 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5281 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5341 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5401 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5461 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5521 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5581 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5641 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5701 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5761 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5821 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5881 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5941 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6001 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6061 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6121 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6181 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6241 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6301 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     6361 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     6421 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     6481 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cattcgacgc
     6541 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     6601 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     6661 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     6721 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     6781 atgccggcca cgatgcgtcc ggcgtagagg atctggctag cgatgaccct gctgattggt
     6841 tcgctgacca tttccgggtg cgggacggcg ttaccagaaa ctcagaaggt tcgtccaacc
     6901 aaaccgactc tgacggcagt ttacgagaga gatgataggg tctgcttcag taagccagat
     6961 gctacacaat taggcttgta catattgtcg ttagaacgcg gctacaatta atacataacc
     7021 ttatgtatca tacacatacg atttaggtga cactatagaa tacacggaat taattc
//
