LOCUS       Exported                7410 bp ds-DNA     circular SYN 02-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7410)
  AUTHORS   Adesnik Lab
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 2024 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7410
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    378..509
                     /label=5'ITR
     misc_feature    570..1041
                     /label=hSyn
     misc_feature    1059..1064
                     /label=BamHI
     misc_feature    1083..1115
                     /label=lox2722
     misc_feature    1167..1199
                     /label=loxP
     primer_bind     1190..1231
                     /label=222_R
     primer_bind     1190..1229
                     /label=S273A_AAV_R
     primer_bind     1192..1231
                     /label=255_F1
     misc_feature    1192..1199
                     /label=lox2722(1)
     CDS             1215..2189
                     /codon_start=1
                     /label=Chronos
                     /translation="METAATMTHAFISAVPSAEATIRGLLSAAAVVTPAADAHGETSNA
                     TTAGADHGCFPHINHGTELQHKIAVGLQWFTVIVAIVQLIFYGWHSFKATTGWEEVYVC
                     VIELVKCFIELFHDVDSPATVYQTNGGAVAWLRYSEWLLTCPVILIHLSNLTGLHEEYS
                     KRTMTILVTDIGNIVWGITAAFTKGPLKILFFMIGLFYGVTCFFQIAKVYIESYHTLPK
                     GVCRKICKIMAYVFFCSWLMFPVMFIAGHEGLGLITPYTSGIGHLILDLIAKNTWGFLG
                     HHLRVKIHEHILIHGDIRKTTTINVAGENMEIETFVDEEEEGGV"
     misc_feature    1443..1460
                     /label=seq primer 3
     misc_feature    2190..2204
                     /label=Linker
     CDS             2205..2228
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2229..2423
                     /codon_start=1
                     /label=Soma-targeting motif
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRF"
     CDS             2439..2495
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(2478..2501)
                     /label=254_F2
     primer_bind     2484..2525
                     /label=254_R1
     CDS             2502..3770
                     /codon_start=1
                     /label=GCaMP8s
                     /translation="MHHHHHHTRRKKTFKEVATAVKIIAMLMGLKINVYIKADKQKNGI
                     KANFHIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSVESKLSKDPNEKRDHMVL
                     LEFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGE
                     GDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYI
                     QERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNLPDQLTEE
                     QIAEYKEAFSLFDKDGDGTITTKELGTVMRSLGHNPTEAELQDMINEVDADGDGTIDFP
                     EFLTMMARKMKYRDTEEEIREAFGVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMI
                     READIDGDGQVNYEEFVQMMTAK"
     CDS             2505..2522
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     2804..2825
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2931..2952)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3192..3211)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(3741..3787)
                     /label=255_R2
     misc_feature    3778..3809
                     /label=lox2722(1)
     misc_feature    3861..3893
                     /label=loxP(1)
     misc_feature    3907..3912
                     /label=HindIII
     misc_feature    3913..4517
                     /label=WPRE
     primer_bind     complement(3972..3992)
                     /label=WPRE-R
     primer_bind     3972..3992
                     /label=WPRE-F
     misc_feature    4556..4768
                     /label=SV40 polyA
     misc_feature    4839..4969
                     /label=3' ITR
     misc_feature    5151..5468
                     /label=f1 origin
     primer_bind     5416..5434
                     /label=f1 ori
     misc_feature    5732..6591
                     /label=Amp
     primer_bind     5884..5903
                     /label=seq primer 7
     primer_bind     6531..6550
                     /label=seq primer 8
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      541 atctacgtag ccatgctcta ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg
      601 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccacccac
      661 tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg
      721 gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt
      781 cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac
      841 tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg
      901 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
      961 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
     1021 cgtgcctgag agcgcagtcg aattcaagct gatagcaagg atcctctaga gtcgactccg
     1081 gaataacttc gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc
     1141 tgctattagc aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat
     1201 ggcgcgccgc cagtatggaa acagccgcca caatgaccca cgcctttatc tcagccgtgc
     1261 ctagcgccga agccacaatt agaggcctgc tgagcgccgc agcagtggtg acaccagcag
     1321 cagacgctca cggagaaacc tctaacgcca caacagccgg agccgatcac ggttgcttcc
     1381 cccacatcaa ccacggaacc gagctgcagc acaagatcgc agtgggactc cagtggttca
     1441 ccgtgatcgt ggctatcgtg cagctcatct tctacggttg gcacagcttc aaggccacaa
     1501 ccggctggga ggaggtctac gtctgcgtga tcgagctcgt caagtgcttc atcgagctgt
     1561 tccacgatgt cgacagccca gccacagtgt accagaccaa cggaggagcc gtggcttggc
     1621 tgcggtacag cgagtggctc ctgacttgcc ccgtgatcct gatccacctg agcaacctga
     1681 ccggactgca cgaagagtac agcaagcgga ccatgaccat cctggtgacc gacatcggca
     1741 acatcgtgtg ggggatcaca gccgccttta caaagggccc cctgaagatc ctgttcttca
     1801 tgatcggcct gttctacggc gtgacttgct tcttccagat cgccaaggtg tatatcgaga
     1861 gctaccacac cctgcccaaa ggcgtctgcc ggaagatttg caagatcatg gcctacgtct
     1921 tcttctgctc ttggctgatg ttccccgtga tgttcatcgc cggacacgag ggactgggcc
     1981 tgatcacacc ttacaccagc ggaatcggcc acctgatcct ggatctgatc gccaagaaca
     2041 cttggggctt cctgggccac cacctgagag tgaagatcca cgagcacatc ctgatccacg
     2101 gcgacatccg gaagacaacc accatcaacg tggccggcga gaacatggag atcgagacct
     2161 tcgtcgacga ggaggaggag ggaggagtgg cggccgcacc ggtagattac aaagacgatg
     2221 acgataagca gtcccagccc atcctcaaca ccaaggagat ggccccgcag agcaagcctc
     2281 cagaggagct ggagatgagc agcatgccca gccccgtggc ccctctgccc gcacgcacgg
     2341 agggcgtcat cgacatgcgg agcatgtcca gcattgacag cttcatcagc tgtgccacgg
     2401 acttccctga agccaccaga ttcgcggcag gaagcggagc tactaacttc agcctgctga
     2461 agcaggctgg agacgtggag gagaaccctg gacctactag tatgcatcat caccatcatc
     2521 acacgcgtcg caagaagacc ttcaaggagg tggccaccgc cgtgaagatc atcgccatgc
     2581 tgatgggcct gaagatcaac gtctatatca aggccgacaa gcagaagaac ggcatcaagg
     2641 cgaacttcca catccgccac aacatcgagg acggcggcgt gcagctcgcc taccactacc
     2701 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcg
     2761 tggagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2821 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggtaccggag
     2881 ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2941 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca
     3001 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3061 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3121 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag gagcgcacca
     3181 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3241 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3301 ggcacaagct ggagtacaac ctgccggacc aactgactga agagcagatc gcagaataca
     3361 aagaggcttt ctccctattt gacaaggacg gggatgggac aataacaacc aaggagctgg
     3421 ggacggtgat gcggtctctc ggacacaacc ccacagaagc agagctgcag gacatgatca
     3481 atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg acaatgatgg
     3541 caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc ggtgtgtttg
     3601 ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg acaaaccttg
     3661 gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac atcgatgggg
     3721 atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagtaa gctagcataa
     3781 cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat
     3841 tctttagaat ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc
     3901 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3961 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     4021 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     4081 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     4141 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4201 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4261 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     4321 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     4381 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4441 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4501 tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga catgataaga
     4561 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     4621 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     4681 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     4741 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     4801 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     4861 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4921 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc
     4981 taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     5041 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     5101 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     5161 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5221 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5281 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5341 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5401 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5461 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5521 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5581 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5641 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5701 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5761 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5821 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5881 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5941 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6001 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6061 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6121 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6181 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6241 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6301 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6361 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6421 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6481 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6541 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6601 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6661 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6721 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6781 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6841 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6901 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6961 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7021 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7081 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7141 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7201 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7261 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7321 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7381 ctggcctttt gctggccttt tgctcacatg
//
