LOCUS       Exported                7148 bp DNA     circular SYN 07-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7148)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 22, 2021 from SnapGene Viewer 5.3.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7148
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          860..999
                     /label=in silico pENTR_L4_L3.seq
     source          1257..1260
                     /label=AGCT inserted
                     /note="AGCT inserted"
     misc_feature    0
                     /label=3' linker
     misc_feature    0
                     /label=5' linker
     misc_feature    1..183
                     /note="ITR"
     primer_bind     382..406
                     /label=pAAV for O1127 - GGTGAACTTCAAGATCCGCCACAAC
     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        517..535
                     /label=GGAATG direct repeat with 3bp spacer
                     /note="GGAATG direct repeat with 3bp spacer"
     enhancer        520..525
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        529..534
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        549..567
                     /label=GGAATG direct repeat with 3bp spacer
                     /note="GGAATG direct repeat with 3bp spacer"
     enhancer        552..557
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        561..566
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        581..599
                     /label=GGAATG direct repeat with 3bp spacer
                     /note="GGAATG direct repeat with 3bp spacer"
     enhancer        584..589
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        593..598
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     misc_feature    605..625
                     /label=spacer 1
                     /note="spacer 1"
     enhancer        634..652
                     /label=GGAATG direct repeat with 3bp spacer
                     /note="GGAATG direct repeat with 3bp spacer"
     enhancer        637..642
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        646..651
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        666..684
                     /label=GGAATG direct repeat with 3bp spacer
                     /note="GGAATG direct repeat with 3bp spacer"
     enhancer        669..674
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        678..683
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        698..716
                     /label=GGAATG direct repeat with 3bp spacer
                     /note="GGAATG direct repeat with 3bp spacer"
     enhancer        701..706
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        710..715
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     misc_feature    722..742
                     /label=spacer 2
                     /note="spacer 2"
     enhancer        751..769
                     /label=GGAATG direct repeat with 3bp spacer
                     /note="GGAATG direct repeat with 3bp spacer"
     enhancer        754..759
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        763..768
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        783..801
                     /label=GGAATG direct repeat with 3bp spacer
                     /note="GGAATG direct repeat with 3bp spacer"
     enhancer        786..791
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        795..800
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        815..833
                     /label=GGAATG direct repeat with 3bp spacer
                     /note="GGAATG direct repeat with 3bp spacer"
     enhancer        818..823
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     enhancer        827..832
                     /label=TEAD binding (GGAATG)
                     /note="TEAD binding (GGAATG)"
     protein_bind    complement(839..859)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        910..950
                     /label=Adenovirus major late promoter
                     /note="Adenovirus major late promoter"
     protein_bind    1000..1020
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1047..1066
                     /label=T3
                     /note="T3"
     promoter        1047..1065
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    1080..1128
                     /label=E01356
                     /note="E01356"
     promoter        1129..1147
                     /label=T7
                     /note="T7"
     promoter        complement(1129..1147)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(1171..1195)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1265..2938
                     /label=luc2
     regulatory      1274..1283
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1280..2932
                     /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"
     primer_bind     complement(1288..1313)
                     /label=O1041
     misc_feature    1390
                     /note="modified base, silent mutation, C-->T"
     misc_feature    2758
                     /note="modified base, point mutation, atg --> atA (Met --> 
                     Ile)"
     misc_feature    2796
                     /note="modified base, point mutation, gtt --> gCt (Val --> 
                     Ala)"
     misc_feature    2954..3546
                     /note="WPRE"
     misc_feature    2960..3548
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3431..3442)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    3565..3833
                     /label=transcription terminator
                     /note="bGH_PA_terminator"
     polyA_signal    3581..3805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3843..4025
                     /note="ITR"
     promoter        complement(4026..4355)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4040..4175
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4542..5175
                     /direction=RIGHT
                     /label=origin of replication
                     /note="pUC"
     rep_origin      complement(4601..5189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5360..6220)
                     /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(6221..6325)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7113..7131
                     /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 agcaggctaa gcttgcagtg gaatgtgtgg aatgtgtctc
      541 aagcttgcag tggaatgtgt ggaatgtgtc tcaagcttgc agtggaatgt gtggaatgtg
      601 tctcacgcgt ggcctcgcct gcaggaagct tgcagtggaa tgtgtggaat gtgtctcaag
      661 cttgcagtgg aatgtgtgga atgtgtctca agcttgcagt ggaatgtgtg gaatgtgtct
      721 cggcctcctg caggaggcgg ccaagcttgc agtggaatgt gtggaatgtg tctcaagctt
      781 gcagtggaat gtgtggaatg tgtctcaagc ttgcagtgga atgtgtggaa tgtgtctcca
      841 acttttctat acaaagttgg caagtgcagg tgccagaaca tttctctggc ctaactggaa
      901 gacgctagcg gggggctata aaagggggtg ggggcgttcg tcctcactct agatctgcga
      961 tctaagtaag cttggcaatc cggtactgtt ggtaaagccc aactttgtat aataaagttg
     1021 agctagttgc taagtctgcc gagtagaatt aaccctcact aaagggtagg tgacactatt
     1081 ctattacatc aatcatctaa gtgcgtcttc tattacttac ctttaatccc tatagtgagt
     1141 cgtattagta gacgcgagtc aatgcatgta cgacagcttt cttgtacaaa gtggtgatat
     1201 cgaattcctg cagcccgggg gatccgcccg ggctagagcg gccgccaccg cggtggagct
     1261 cgcgactagt gacgccacca tggaagatgc caaaaacatt aagaagggcc cagcgccatt
     1321 ctacccactc gaagacggga ccgccggcga gcagctgcac aaagccatga agcgctacgc
     1381 cctggtgcct ggcaccatcg cctttaccga cgcacatatc gaggtggaca ttacctacgc
     1441 cgagtacttc gagatgagcg ttcggctggc agaagctatg aagcgctatg ggctgaatac
     1501 aaaccatcgg atcgtggtgt gcagcgagaa tagcttgcag ttcttcatgc ccgtgttggg
     1561 tgccctgttc atcggtgtgg ctgtggcccc agctaacgac atctacaacg agcgcgagct
     1621 gctgaacagc atgggcatca gccagcccac cgtcgtattc gtgagcaaga aagggctgca
     1681 aaagatcctc aacgtgcaaa agaagctacc gatcatacaa aagatcatca tcatggatag
     1741 caagaccgac taccagggct tccaaagcat gtacaccttc gtgacttccc atttgccacc
     1801 cggcttcaac gagtacgact tcgtgcccga gagcttcgac cgggacaaaa ccatcgccct
     1861 gatcatgaac agtagtggca gtaccggatt gcccaagggc gtagccctac cgcaccgcac
     1921 cgcttgtgtc cgattcagtc atgcccgcga ccccatcttc ggcaaccaga tcatccccga
     1981 caccgctatc ctcagcgtgg tgccatttca ccacggcttc ggcatgttca ccacgctggg
     2041 ctacttgatc tgcggctttc gggtcgtgct catgtaccgc ttcgaggagg agctattctt
     2101 gcgcagcttg caagactata agattcaatc tgccctgctg gtgcccacac tatttagctt
     2161 cttcgctaag agcactctca tcgacaagta cgacctaagc aacttgcacg agatcgccag
     2221 cggcggggcg ccgctcagca aggaggtagg tgaggccgtg gccaaacgct tccacctacc
     2281 aggcatccgc cagggctacg gcctgacaga aacaaccagc gccattctga tcacccccga
     2341 aggggacgac aagcctggcg cagtaggcaa ggtggtgccc ttcttcgagg ctaaggtggt
     2401 ggacttggac accggtaaga cactgggtgt gaaccagcgc ggcgagctgt gcgtccgtgg
     2461 ccccatgatc atgagcggct acgttaacaa ccccgaggct acaaacgctc tcatcgacaa
     2521 ggacggctgg ctgcacagcg gcgacatcgc ctactgggac gaggacgagc acttcttcat
     2581 cgtggaccgg ctgaagagcc tgatcaaata caagggctac caggtagccc cagccgaact
     2641 ggagagcatc ctgctgcaac accccaacat cttcgacgcc ggggtcgccg gcctgcccga
     2701 cgacgatgcc ggcgagctgc ccgccgcagt cgtcgtgctg gaacacggta aaaccataac
     2761 cgagaaggag atcgtggact atgtggccag ccaggctaca accgccaaga agctgcgcgg
     2821 tggtgttgtg ttcgtggacg aggtgcctaa aggactgacc ggcaagttgg acgcccgcaa
     2881 gatccgcgag attctcatta aggccaagaa gggcggcaag atcgccgtgt aattcgaatt
     2941 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3001 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3061 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3121 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3181 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3241 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3301 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3361 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3421 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3481 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3541 ctccccgcat cgataccgtc gactcgctga tcagcctcga ctgtgccttc tagttgccag
     3601 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3661 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3721 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3781 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgact
     3841 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     3901 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3961 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4021 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     4081 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     4141 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
     4201 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     4261 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     4321 gagcctgggg actttccaca ccctaactga cacacattcc acagctgcat taatgaatcg
     4381 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4441 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4501 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4561 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4621 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4681 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4741 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4801 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4861 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4921 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4981 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5041 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5101 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5161 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5221 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5281 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5341 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5401 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5461 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5521 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5581 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5641 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5701 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5761 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5821 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5881 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5941 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6001 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6061 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6121 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6181 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6241 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6301 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     6361 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     6421 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     6481 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     6541 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     6601 accattcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     6661 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     6721 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     6781 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     6841 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctggct agcgatgacc
     6901 ctgctgattg gttcgctgac catttccggg tgcgggacgg cgttaccaga aactcagaag
     6961 gttcgtccaa ccaaaccgac tctgacggca gtttacgaga gagatgatag ggtctgcttc
     7021 agtaagccag atgctacaca attaggcttg tacatattgt cgttagaacg cggctacaat
     7081 taatacataa ccttatgtat catacacata cgatttaggt gacactatag aatacacgga
     7141 attaattc
//
