LOCUS       AAV-PB-SRT-tdTom        6472 bp DNA     circular SYN 10-OCT-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6472)
  AUTHORS   Jeffrey Milbrandt Lab WUSTL
  TITLE     Direct Submission
  JOURNAL   Exported Oct 10, 2022 from SnapGene 6.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6472
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     terminator      1..44
                     /label=tR2 lambda phage terminator
     primer_bind     392..409
                     /label=M13-21
     primer_bind     394..410
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        421..439
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    448..456
                     /label=vector ID
     misc_feature    457..588
                     /label=5'ITR
                     /note="5'ITR"
     misc_feature    615..716
                     /label=CAG Enhancer
                     /note="CAG Enhancer"
     primer_bind     701..736
                     /label=AAVSRTF4
     LTR             complement(735..934)
                     /label=pBac 3'-LTR
     promoter        1024..2221
                     /label=EF1a Promoter
                     /label=EF1a\Promoter
     misc_feature    1024..1033
                     /label=removable
     promoter        1040..2221
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     misc_feature    1237..1269
                     /label=exon1
     intron          1270..2212
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human
                     EF-1-alpha"
     regulatory      2229..2238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2235..3665
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    2235..2237
                     /label=PAC
                     /note="PuroR"
     primer_bind     2971..2991
                     /label=SRT_tdt_F2
     primer_bind     3634..3653
                     /label=SRT_tdTomato_F1
     misc_feature    complement(3681..3690)
                     /label=bar02
     misc_feature    complement(3691..3727)
                     /label=IllumiaSeq2
     misc_feature    complement(3728..3735)
                     /label=Pme1
     misc_feature    complement(3752..3759)
                     /label=Mre
     LTR             3769..4074
                     /label=pBac 5'-LTR
     misc_feature    4069..4072
                     /label=Barcode CCAA
     misc_feature    4075..4078
                     /label=TTAA
     misc_feature    4091..4173
                     /label=N117 Sm1 Rz
                     /note="Hammerhead ribozyme N117"
     misc_feature    4199..4305
                     /label=5S ribosomal RNA
     misc_feature    4306..4317
                     /label=5S ribosomal RNA
     primer_bind     complement(4325..4358)
                     /label=AAVSRTR4
     misc_recomb     4388..4518
                     /label=3' ITR
                     /note="3' ITR"
     misc_feature    4528..4536
                     /label=vector ID
     primer_bind     complement(4590..4608)
                     /label=AAVSRTR4
     terminator      complement(4683..4726)
                     /label=rrnB T1 transcription terminator
     rep_origin      complement(4783..5371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5378..5386
                     /label=TCF/LEF binding
                     /bound_moiety="TCF LEF"
     CDS             complement(5542..6402)
                     /codon_start=1
                     /label=ampR - bla
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6403..6472)
                     /label=bla promoter
ORIGIN
        1 gctaataaca ggcctgctgg taatcgcagg cctttttatt tggtgtattt agaaaaataa
       61 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
      121 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
      181 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtggccgc
      241 tacagggcgc tcccattcgc cattcaggct gcgcaactgt tgggaagggc gtttcggtgc
      301 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      361 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgacg
      421 taatacgact cactataggg cgaattgtag gcggaaggct gcgcgctcgc tcgctcactg
      481 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
      541 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctat tgacgtcaga
      601 tcgctaaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      661 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatacgtc
      721 aaggccgcat ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg
      781 tgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa
      841 tagatattaa gttttattat atttacactt acatactaat aataaattca acaaacaatt
      901 tatttatgtt tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa
      961 caaaactttt atcgaattcc tgcagcccag cttgatctta ggtccctcga cgaattctct
     1021 agagaagagc atgcgtgagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     1081 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc
     1141 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     1201 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     1261 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc
     1321 ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc ttgatcccga
     1381 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
     1441 tcgtgcttga gttgaggcct ggcttgggcg ctggggccgc cgcgtgcgaa tctggtggca
     1501 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
     1561 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
     1621 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     1681 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     1741 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     1801 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     1861 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     1921 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     1981 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     2041 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     2101 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     2161 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     2221 aaaactacgc caccatggtg agcaagggcg aggaggtcat caaagagttc atgcgcttca
     2281 aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg
     2341 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
     2401 ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc
     2461 accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc
     2521 gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg
     2581 acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg
     2641 taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg
     2701 gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg
     2761 tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg
     2821 tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2881 agcgctccga gggccgccac cacctgttcc tggggcatgg caccggcagc accggcagcg
     2941 gcagctccgg caccgcctcc tccgaggaca acaacatggc cgtcatcaaa gagttcatgc
     3001 gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg
     3061 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc
     3121 ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg
     3181 tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt
     3241 gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc
     3301 tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg
     3361 gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc
     3421 gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact
     3481 acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact
     3541 actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac
     3601 agtacgagcg ctccgagggc cgccaccacc tgttcctgta cggcatggac gagctgtaca
     3661 agtaacaact cgagtaccgg cttaagtgat agatcggaag agcggttcag caggaatgcc
     3721 gagaccggtt taaactcagt gtccatagcg acgccggcgc gataagctat ctataacaag
     3781 aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg
     3841 tatgagttaa atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg
     3901 tcgttatagt tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc
     3961 caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga
     4021 gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttcc aaggttaaga
     4081 gctcaagctt ctgagatgca ggtacatccc actgatgagt cccaaatagg acgaaacgcg
     4141 cttcggtgcg tctgggattc cactgctatc cacgcggccg cacggcatta cgttagacgt
     4201 ctaggccaca ccaccctgaa ggcgcctgct cgcctctgat ctgttgaagc taagcagggt
     4261 cggtcctggt tagtacttgg atgggactcc gcctggtaat agccggtacc gtaggcttgc
     4321 atggtcatag ctgtttcctt gcgctgggga tgcggtgggc tctatagaga gtcgatcgtg
     4381 tgatcgaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4441 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4501 agcgagcgag cgcgcagcat gtagagattc cgcctatcac tgcccgcttt ccagtcggga
     4561 aacctgtcgt gccagctgca ttaacatggt catagctgtt tccttgcgta ttgggcgctc
     4621 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc gggtaaagcc tggggtgcct
     4681 aaataaaacg aaaggcccag tcttccgact gagcctttcg ttttattgag caaaaggcca
     4741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5581 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5641 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc
     5701 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5761 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5821 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5881 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5941 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6001 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6061 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6121 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6181 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6241 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6301 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6361 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6421 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aa
//
