LOCUS       Exported                7218 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CAG-RFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7218)
  AUTHORS   Nathanson JL, Yanagawa Y, Obata K, Callaway EM
  TITLE     Preferential labeling of inhibitory and excitatory cortical neurons 
            by endogenous tropism of adeno-associated virus and lentivirus 
            vectors.
  JOURNAL   Neuroscience. 2009 Jun 30. 161(2):441-50.
  PUBMED    19318117
REFERENCE   2  (bases 1 to 7218)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7218
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     intron          27..1043
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     967..989
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1051..1070
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1154..1629
                     /label=-beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     primer_bind     1711..1727
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1711..1727
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      1760..1769
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1766..2443
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(1896..1914)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1946..1966)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2164..2183
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2358..2381
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        complement(2485..2503)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2486..2503)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2523..2540)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2529..2640
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2784..2913
                     /label=AAV2 ITR
     rep_origin      3284..3797
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3371..3390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3581..3602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4279..4298)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4398..4420
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4458..4476)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4544..4648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4649..5509
                     /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(4867..4886)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5680..6268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6169..6188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6422..6439
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        6586..6965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6967..26
                     /label=chicken beta-actin promoter
ORIGIN
        1 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
       61 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
      121 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc
      181 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg
      241 gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct
      301 ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca
      361 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg
      421 ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt
      481 cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt
      541 cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
      601 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg
      661 ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc
      721 ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc
      781 cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg
      841 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
      901 tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca
      961 gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc
     1021 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc
     1081 attttggcaa agaattgtgg cggcccgacc gcgggctagc ggcgcgccga gctcggatcc
     1141 tgagaacttc agggtgagtc tatgggaccc ttgatgtttt ctttcccctt cttttctatg
     1201 gttaagttca tgtcatagga aggggagaag taacagggta cacatattga ccaaatcagg
     1261 gtaattttgc atttgtaatt ttaaaaaatg ctttcttctt ttaatatact tttttgttta
     1321 tcttatttct aatactttcc ctaatctctt tctttcaggg caataatgat acaatgtatc
     1381 atgcctcttt gcaccattct aaagaataac agtgataatt tctgggttaa ggcaatagca
     1441 atatttctgc atataaatat ttctgcatat aaattgtaac tgatgtaaga ggtttcatat
     1501 tgctaatagc agctacaatc cagctaccat tctgctttta ttttatggtt gggataaggc
     1561 tggattattc tgagtccaag ctaggccctt ttgctaatca tgttcatacc tcttatcttc
     1621 ctcccacagc tcctgggcaa cgtgctggtc tgtgtgctgg cccatcactt tggcaaagca
     1681 cgtgaattcg aagatctggc cgcctcggcc tctagaacta gtggatcccc cgggctgcag
     1741 gaattcgatg ctaccggtcg ccaccatggc ctcctccgag gacgtcatca aggagttcat
     1801 gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     1861 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     1921 ccccctgccc ttcgcctggg acatcctgtc cccccagttc cagtacggct ccaaggtgta
     1981 cgtgaagcac cccgccgaca tccccgacta caagaagctg tccttccccg agggcttcaa
     2041 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     2101 cctgcaggac ggctccttca tctacaaggt gaagttcatc ggcgtgaact tcccctccga
     2161 cggccccgta atgcagaaga agactatggg ctgggaggcc tccaccgagc gcctgtaccc
     2221 ccgcgacggc gtgctgaagg gcgagatcca caaggccctg aagctgaagg acggcggcca
     2281 ctacctggtg gagttcaagt ccatctacat ggccaagaag cccgtgcagc tgcccggcta
     2341 ctactacgtg gactccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     2401 gcagtacgag cgcgccgagg gccgccacca cctgttcctg tagcggccaa atgaattccc
     2461 aaaccgtacg ggaaacggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     2521 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2581 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2641 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2701 ggaggattgg gaagacaata gcgcggccgc tctagagcat ggctacgtag ataagtagca
     2761 tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct
     2821 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     2881 ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag aggcccgcac
     2941 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgattcc gttgcaatgg
     3001 ctggcggtaa tattgttctg gatattacca gcaaggccga tagtttgagt tcttctactc
     3061 aggcaagtga tgttattact aatcaaagaa gtattgcgac aacggttaat ttgcgtgatg
     3121 gacagactct tttactcggt ggcctcactg attataaaaa cacttctcag gattctggcg
     3181 taccgttcct gtctaaaatc cctttaatcg gcctcctgtt tagctcccgc tctgattcta
     3241 acgaggaaag cacgttatac gtgctcgtca aagcaaccat agtacgcgcc ctgtagcggc
     3301 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3361 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3421 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3481 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3541 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3601 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3661 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3721 atattaacgc ttacaattta aatatttgct tatacaatct tcctgttttt ggggcttttc
     3781 tgattatcaa ccggggtaca tatgattgac atgctagttt tacgattacc gttcatcgat
     3841 tctcttgttt gctccagact ctcaggcaat gacctgatag cctttgtaga gacctctcaa
     3901 aaatagctac cctctccggc atgaatttat cagctagaac ggttgaatat catattgatg
     3961 gtgatttgac tgtctccggc ctttctcacc cgtttgaatc tttacctaca cattactcag
     4021 gcattgcatt taaaatatat gagggttcta aaaattttta tccttgcgtt gaaataaagg
     4081 cttctcccgc aaaagtatta cagggtcata atgtttttgg tacaaccgat ttagctttat
     4141 gctctgaggc tttattgctt aattttgcta attctttgcc ttgcctgtat gatttattgg
     4201 atgttggaat cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4261 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4321 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4381 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4441 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4501 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4561 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4621 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4681 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4741 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4801 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4861 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4921 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4981 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5041 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5101 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5161 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5221 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5281 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5341 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5401 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5461 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5521 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5581 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5641 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5701 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5761 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5821 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5881 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5941 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6001 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6061 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6121 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6181 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6241 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6301 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6361 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6421 cagcgagtca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     6481 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctagagg
     6541 atccttcgaa aagcttgtat acggccggcc acgcgttcgc cactcgacat tgattattga
     6601 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6661 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6721 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6781 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6841 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6901 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6961 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
     7021 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
     7081 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
     7141 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     7201 ggcggcggcg gcggccct
//
