LOCUS       Exported                7027 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    AAV-EF1a-BbTagBY
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7027)
  AUTHORS   Cai D, Cohen KB, Luo T, Lichtman JW, Sanes JR
  TITLE     Improved tools for the Brainbow toolbox.
  JOURNAL   Nat Methods. 2013 May 5;10(6):540-7. doi: 10.1038/nmeth.2450. Epub 
            2013 May 5.
  PUBMED    23817127
REFERENCE   2  (bases 1 to 7027)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7027
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1416..1449
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      2220..2229
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(2230..2263)
                     /label=loxKR3
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    2267..2300
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(2379..3095)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2421..2442)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2770..2789
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3029..3050
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3092..3101
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(3102..3135)
                     /label=loxKR3
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3154..3742
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3207..3227)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3625..3636)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3774..4250
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3973..3992)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4290..4430
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4290..4419
                     /label=AAV2 ITR
     rep_origin      4505..4960
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4592..4611)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4802..4823
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4977..4996)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5096..5118
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5156..5174)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5242..5346
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5347..6207
                     /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"
     primer_bind     complement(5565..5584)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6378..6966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6867..6886
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtacctaccg ttcgtatagc atacattata
     1441 cgaagttatt taggagagca cacacttgca gctcatgcag ccggggccac tctcatcagg
     1501 agggttcagc ttactcacgc gtgcacttct gtgccccagt ttgctaggga ggtcgcagta
     1561 tctggccact gccacctcgt gctgctcgac gtaggtctcg ttgttggcct ccttgattct
     1621 ttccagtctg tagtccacat agtagacgcc aggcatcttg aggttcttag cgggtttctt
     1681 ggatctatat gtggtcttga tgtttgcgat cagatggctc ccgcccacga gcttcagggc
     1741 catgtcgttt ctgccttcca ggccgccgtc agcggggtac agcgtctcgg tgaaggcctc
     1801 ccagccgagt gttttcttct gcatcacagg gccgttggat gtgaagttca cccctctgat
     1861 cttgacgttg tagatgaggc agccgtcctg gaggctggtg tcctgggtag cggtcagcac
     1921 gcccccgtct tcgtatgtgg tgactctctc ccatgtgaag ccctcaggga aggactgctt
     1981 gaagaagtcg gggatgccct gggtgtggtt gatgaaggtc ttgctgccgt agaggaagct
     2041 agtagccagg atgtcgaagg cgaaggggag agggccgccc tcgaccacct tgattctcat
     2101 ggtctgggtg ccctcgtagg gcttgccttc gccctcggat gtgcacttga agtgatggtt
     2161 gtccacggtg ccctccatgt acagcttcat gtgcatgttc tccttaatca gctcgctcac
     2221 catggtggca taacaaggta taatgtatgc tatacgaagt tatgatataa cttcgtatag
     2281 catacattat acgaacggta tcaggagagc acacacttgc agctcatgca gccggggcca
     2341 ctctcatcag gagggttcag cttactcacg cgtgcactct tgtacagctc gtccatgccg
     2401 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     2461 gggtctttgc tcagggcgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg
     2521 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
     2581 atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata
     2641 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     2701 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     2761 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     2821 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg
     2881 aagcactgca ggccgtagcc gaaggtggtc acgagggtgg gccagggcac gggcagcttg
     2941 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     3001 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     3061 accccggtga acagctcctc gcccttgctc accatggtgg cataacaagg tataatgtat
     3121 gctatacgaa gttatgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     3181 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3241 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3301 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3361 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3421 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3481 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3541 tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc
     3601 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3661 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3721 tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc
     3781 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     3841 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     3901 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     3961 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     4021 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     4081 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     4141 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     4201 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt
     4261 aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     4321 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4381 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     4441 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     4501 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4561 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     4621 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4681 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4741 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4801 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     4861 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     4921 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     4981 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     5041 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     5101 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     5161 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5221 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5281 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5341 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5401 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5461 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5521 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5581 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5641 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5701 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5761 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5821 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5881 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5941 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6001 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6061 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6121 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6181 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6241 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6301 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6361 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6421 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6481 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6541 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6601 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6661 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6721 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6781 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6841 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6901 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6961 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7021 cacatgt
//
