LOCUS       Exported                7031 bp ds-DNA     circular SYN 01-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7031)
  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..7031
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    378..509
                     /label=5'ITR
     misc_feature    570..1041
                     /label=hSyn
     primer_bind     1048..1084
                     /label=242_F
     misc_feature    1059..1063
                     /label=BamHI
     CDS             1071..2045
                     /codon_start=1
                     /label=Chronos
                     /translation="METAATMTHAFISAVPSAEATIRGLLSAAAVVTPAADAHGETSNA
                     TTAGADHGCFPHINHGTELQHKIAVGLQWFTVIVAIVQLIFYGWHSFKATTGWEEVYVC
                     VIELVKCFIELFHDVDSPATVYQTNGGAVAWLRYSEWLLTCPVILIHLSNLTGLHEEYS
                     KRTMTILVTDIGNIVWGITAAFTKGPLKILFFMIGLFYGVTCFFQIAKVYIESYHTLPK
                     GVCRKICKIMAYVFFCSWLMFPVMFIAGHEGLGLITPYTSGIGHLILDLIAKNTWGFLG
                     HHLRVKIHEHILIHGDIRKTTTINVAGENMEIETFVDEEEEGGV"
     misc_feature    1299..1316
                     /label=seq primer 3
     misc_feature    1490
                     /label=M-E:GAG
     primer_bind     complement(1586..1626)
                     /label=NG_GA_R1
     primer_bind     1590..1627
                     /label=NG_GA_F2
     misc_feature    1923..1939
                     /label=seq primer 2
     primer_bind     complement(2041..2071)
                     /label=243_R1
     misc_feature    2046..2053
                     /label=Linker
     CDS             2055..3317
                     /codon_start=1
                     /label=GCaMP8m
                     /translation="HHHHHHTRRKKTFKEVATAVKIIGRLMGLKINVYIKADKQKNGIK
                     ANFHIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSVESKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEG
                     DATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQ
                     ERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNLPDQLTEEQ
                     IAEYKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGDGTIDFPE
                     FLTMMARKMKYRDTEEEIREAFGVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIR
                     EADIDGDGQVNYEEFVQMMTAK"
     CDS             2055..2072
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     2354..2375
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2481..2502)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2742..2761)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3323
                     /label=Linker
     misc_feature    3324..3521
                     /label=Soma-targeting motif
     primer_bind     complement(3497..3538)
                     /label=242_R
     misc_feature    3528..3533
                     /label=HindIII
     misc_feature    3534..4138
                     /label=WPRE
     primer_bind     complement(3593..3613)
                     /label=WPRE-R
     primer_bind     3593..3613
                     /label=WPRE-F
     misc_feature    4177..4389
                     /label=SV40 polyA
     misc_feature    4460..4590
                     /label=3' ITR
     misc_feature    4772..5089
                     /label=f1 origin
     primer_bind     5037..5055
                     /label=f1 ori
     misc_feature    5353..6212
                     /label=Amp
     primer_bind     5505..5524
                     /label=seq primer 7
     primer_bind     6152..6171
                     /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 atccgctagt atggaaacag
     1081 ccgccacaat gacccacgcc tttatctcag ccgtgcctag cgccgaagcc acaattagag
     1141 gcctgctgag cgccgcagca gtggtgacac cagcagcaga cgctcacgga gaaacctcta
     1201 acgccacaac agccggagcc gatcacggtt gcttccccca catcaaccac ggaaccgagc
     1261 tgcagcacaa gatcgcagtg ggactccagt ggttcaccgt gatcgtggct atcgtgcagc
     1321 tcatcttcta cggttggcac agcttcaagg ccacaaccgg ctgggaggag gtctacgtct
     1381 gcgtgatcga gctcgtcaag tgcttcatcg agctgttcca cgatgtcgac agcccagcca
     1441 cagtgtacca gaccaacgga ggagccgtgg cttggctgcg gtacagcgag tggctcctga
     1501 cttgccccgt gatcctgatc cacctgagca acctgaccgg actgcacgaa gagtacagca
     1561 agcggaccat gaccatcctg gtgaccgaca tcggcaacat cgtgtggggg atcacagccg
     1621 cctttacaaa gggccccctg aagatcctgt tcttcatgat cggcctgttc tacggcgtga
     1681 cttgcttctt ccagatcgcc aaggtgtata tcgagagcta ccacaccctg cccaaaggcg
     1741 tctgccggaa gatttgcaag atcatggcct acgtcttctt ctgctcttgg ctgatgttcc
     1801 ccgtgatgtt catcgccgga cacgagggac tgggcctgat cacaccttac accagcggaa
     1861 tcggccacct gatcctggat ctgatcgcca agaacacttg gggcttcctg ggccaccacc
     1921 tgagagtgaa gatccacgag cacatcctga tccacggcga catccggaag acaaccacca
     1981 tcaacgtggc cggcgagaac atggagatcg agaccttcgt cgacgaggag gaggagggag
     2041 gagtggcggc cgcccatcat caccatcatc acacgcgtcg caagaagacc ttcaaggagg
     2101 tggccaccgc cgtgaagatc atcggccgcc tgatgggcct gaagatcaac gtctatatca
     2161 aggccgacaa gcagaagaac ggcatcaagg cgaacttcca catccgccac aacatcgagg
     2221 acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg
     2281 tgctgctgcc cgacaaccac tacctgagcg tggagtccaa actttcgaaa gaccccaacg
     2341 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2401 tggacgagct gtacaagggc ggtaccggag ggagcatggt gagcaagggc gaggagctgt
     2461 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2521 gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2581 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2641 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2701 tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2761 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2821 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ctgccggacc
     2881 aactgactga agagcagatc gcagaataca aagaggcttt ctccctattt gacaaggacg
     2941 gggatgggac aataacaacc aaggagctgg ggacggtgat gcggtctctg gggcagaacc
     3001 ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacaa
     3061 tcgacttccc tgagttcctg acaatgatgg caagaaaaat gaaatacagg gacacggaag
     3121 aagaaattag agaagcgttc ggtgtgtttg ataaggatgg caatggctac atcagtgcag
     3181 cagagcttcg ccacgtgatg acaaaccttg gagagaagtt aacagatgaa gaggttgatg
     3241 aaatgatcag ggaagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtac
     3301 aaatgatgac agcgaagcaa ttgcagtccc agcccatcct caacaccaag gagatggccc
     3361 cgcagagcaa gcctccagag gagctggaga tgagcagcat gcccagcccc gtggcccctc
     3421 tgcccgcacg cacggagggc gtcatcgaca tgcggagcat gtccagcatt gacagcttca
     3481 tcagctgtgc cacggacttc cctgaagcca ccagattcta agatatcaag cttatcgata
     3541 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3601 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3661 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3721 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3781 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3841 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3901 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3961 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4021 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4081 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     4141 gacctcgacc cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     4201 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     4261 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     4321 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     4381 aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc
     4441 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4501 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4561 gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact ggccgtcgtt
     4621 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     4681 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     4741 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4801 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4861 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4921 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4981 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5041 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5101 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5161 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     5221 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5281 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5341 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5401 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5461 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5521 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5581 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5641 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5701 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5761 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5821 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5881 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5941 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6001 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6061 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6121 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6181 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6241 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6301 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6361 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6421 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6481 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6541 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6601 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6661 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6721 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6781 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6841 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6901 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6961 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7021 ttgctcacat g
//
