LOCUS       Exported                7100 bp ds-DNA     circular SYN 01-AUG-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV_ER-RE_MLP_BC1064-luc2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7100)
  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..7100
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          251..1221
                     /label=seq verified pAAV
                     /note="seq verified pAAV"
     source          812..951
                     /label=in silico pENTR_L4_L3.seq
     source          1209..1212
                     /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..588
                     /label=4x ER-RE
                     /note="4x ER-RE"
     enhancer        514..528
                     /label=ERE
                     /note="ERE"
     variation       534..548
                     /label=ERE
                     /note="ERE"
     enhancer        554..568
                     /label=ERE
                     /note="ERE"
     enhancer        574..588
                     /label=ERE
                     /note="ERE"
     misc_feature    589..609
                     /label=spacer 1
                     /note="spacer 1"
     enhancer        610..689
                     /label=4x ER-RE
                     /note="4x ER-RE"
     enhancer        615..629
                     /label=ERE
                     /note="ERE"
     variation       635..649
                     /label=ERE
                     /note="ERE"
     enhancer        655..669
                     /label=ERE
                     /note="ERE"
     enhancer        675..689
                     /label=ERE
                     /note="ERE"
     misc_feature    690..710
                     /label=spacer 2
                     /note="spacer 2"
     enhancer        711..790
                     /label=4x ER-RE
                     /note="4x ER-RE"
     enhancer        716..730
                     /label=ERE
                     /note="ERE"
     variation       736..750
                     /label=ERE
                     /note="ERE"
     enhancer        756..770
                     /label=ERE
                     /note="ERE"
     enhancer        776..790
                     /label=ERE
                     /note="ERE"
     protein_bind    complement(791..811)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        862..902
                     /label=Adenovirus major late promoter
                     /note="Adenovirus major late promoter"
     protein_bind    952..972
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     973..998
                     /label=Dec1 #10243
                     /note="Dec1 #10243"
     promoter        999..1018
                     /label=T3
                     /note="T3"
     promoter        999..1017
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1019..1031
                     /label=truncated SP6
                     /note="truncated SP6"
     misc_feature    1032..1080
                     /label=E01064
     promoter        complement(1081..1099)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1100..1125)
                     /label=Dec2 #9752
                     /note="Dec2 #9752"
     protein_bind    complement(1123..1147)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1226..1235
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1232..2884
                     /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    1342
                     /label=modified base, silent mutation, C--&gt;T
                     /note="modified base, silent mutation, C-->T"
     misc_feature    2710
                     /label=modified base
                     /note="modified base, point mutation, atg --> atA (Met --> 
                     Ile)"
     misc_feature    2748
                     /label=modified base
                     /note="modified base, point mutation, gtt --> gCt (Val --> 
                     Ala)"
     misc_feature    2906..3498
                     /label=WPRE
                     /note="WPRE"
     misc_feature    2912..3500
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3383..3394)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    3517..3785
                     /label=bGH_PA_terminator
                     /note="bGH_PA_terminator"
     polyA_signal    3533..3757
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3795..3977
                     /label=ITR
                     /note="ITR"
     promoter        complement(3978..4307)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        complement(3985..4294)
                     /label=SV40 promotor and origin
                     /note="SV40 promotor and origin"
     rep_origin      3992..4127
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4494..5127
                     /direction=RIGHT
                     /label=pUC
                     /note="pUC"
     rep_origin      complement(4553..5141)
                     /direction=LEFT
                     /label=ori (pUC)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5312..6172)
                     /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(6173..6277)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7065..7083
                     /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 cccaggtcac cctgacctgg cccaggtcat
      541 cctgacctgg cccaggtcat tctgacctgg cccaggtcac cctgacctac gcgtggcctc
      601 gcctgcaggg gcccaggtca ccctgacctg gcccaggtca tcctgacctg gcccaggtca
      661 ttctgacctg gcccaggtca ccctgacctg gcctcctgca ggaggcggcc ggcccaggtc
      721 accctgacct ggcccaggtc atcctgacct ggcccaggtc attctgacct ggcccaggtc
      781 accctgacct caacttttct atacaaagtt ggcaagtgca ggtgccagaa catttctctg
      841 gcctaactgg aagacgctag cggggggcta taaaaggggg tgggggcgtt cgtcctcact
      901 ctagatctgc gatctaagta agcttggcaa tccggtactg ttggtaaagc ccaactttgt
      961 ataataaagt tgagctagtt gctaagtctg ccgagtagaa ttaaccctca ctaaagggta
     1021 ggtgacacta tttactacta catcaaactt ttgacgctga cttttctaaa tcacattact
     1081 cctatagtga gtcgtattag tagacgcgag tcaatgcatg tacgacagct ttcttgtaca
     1141 aagtggtgat atcgaattcc tgcagcccgg gggatccgcc cgggctagag cggccgccac
     1201 cgcggtggag ctcgcgacta gtgacgccac catggaagat gccaaaaaca ttaagaaggg
     1261 cccagcgcca ttctacccac tcgaagacgg gaccgccggc gagcagctgc acaaagccat
     1321 gaagcgctac gccctggtgc ctggcaccat cgcctttacc gacgcacata tcgaggtgga
     1381 cattacctac gccgagtact tcgagatgag cgttcggctg gcagaagcta tgaagcgcta
     1441 tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag aatagcttgc agttcttcat
     1501 gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc ccagctaacg acatctacaa
     1561 cgagcgcgag ctgctgaaca gcatgggcat cagccagccc accgtcgtat tcgtgagcaa
     1621 gaaagggctg caaaagatcc tcaacgtgca aaagaagcta ccgatcatac aaaagatcat
     1681 catcatggat agcaagaccg actaccaggg cttccaaagc atgtacacct tcgtgacttc
     1741 ccatttgcca cccggcttca acgagtacga cttcgtgccc gagagcttcg accgggacaa
     1801 aaccatcgcc ctgatcatga acagtagtgg cagtaccgga ttgcccaagg gcgtagccct
     1861 accgcaccgc accgcttgtg tccgattcag tcatgcccgc gaccccatct tcggcaacca
     1921 gatcatcccc gacaccgcta tcctcagcgt ggtgccattt caccacggct tcggcatgtt
     1981 caccacgctg ggctacttga tctgcggctt tcgggtcgtg ctcatgtacc gcttcgagga
     2041 ggagctattc ttgcgcagct tgcaagacta taagattcaa tctgccctgc tggtgcccac
     2101 actatttagc ttcttcgcta agagcactct catcgacaag tacgacctaa gcaacttgca
     2161 cgagatcgcc agcggcgggg cgccgctcag caaggaggta ggtgaggccg tggccaaacg
     2221 cttccaccta ccaggcatcc gccagggcta cggcctgaca gaaacaacca gcgccattct
     2281 gatcaccccc gaaggggacg acaagcctgg cgcagtaggc aaggtggtgc ccttcttcga
     2341 ggctaaggtg gtggacttgg acaccggtaa gacactgggt gtgaaccagc gcggcgagct
     2401 gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac aaccccgagg ctacaaacgc
     2461 tctcatcgac aaggacggct ggctgcacag cggcgacatc gcctactggg acgaggacga
     2521 gcacttcttc atcgtggacc ggctgaagag cctgatcaaa tacaagggct accaggtagc
     2581 cccagccgaa ctggagagca tcctgctgca acaccccaac atcttcgacg ccggggtcgc
     2641 cggcctgccc gacgacgatg ccggcgagct gcccgccgca gtcgtcgtgc tggaacacgg
     2701 taaaaccata accgagaagg agatcgtgga ctatgtggcc agccaggcta caaccgccaa
     2761 gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct aaaggactga ccggcaagtt
     2821 ggacgcccgc aagatccgcg agattctcat taaggccaag aagggcggca agatcgccgt
     2881 gtaattcgaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     2941 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3001 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3061 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3121 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3181 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3241 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3301 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3361 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3421 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3481 cctttgggcc gcctccccgc atcgataccg tcgactcgct gatcagcctc gactgtgcct
     3541 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3601 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3661 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3721 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3781 tggggctcga ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta
     3841 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3901 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3961 gcgagcgcgc agagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     4021 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     4081 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     4141 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     4201 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     4261 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgc
     4321 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4381 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4441 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4501 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4561 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4621 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4681 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4741 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4801 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4861 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4921 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4981 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5041 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5101 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5161 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5221 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5281 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5341 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5401 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5461 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5521 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5581 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5641 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5701 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5761 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5821 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5881 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5941 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6001 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6061 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6121 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6181 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6241 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6301 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     6361 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     6421 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     6481 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     6541 tactgagagt gcaccattcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     6601 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     6661 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     6721 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     6781 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgg
     6841 ctagcgatga ccctgctgat tggttcgctg accatttccg ggtgcgggac ggcgttacca
     6901 gaaactcaga aggttcgtcc aaccaaaccg actctgacgg cagtttacga gagagatgat
     6961 agggtctgct tcagtaagcc agatgctaca caattaggct tgtacatatt gtcgttagaa
     7021 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
     7081 agaatacacg gaattaattc
//
