LOCUS       Exported                7305 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 7305)
  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..7305
                     /organism="synthetic DNA construct"
                     /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     1192..1228
                     /label=243_R2
     misc_feature    1209..1406
                     /label=Soma-targeting motif
     primer_bind     complement(1386..1429)
                     /label=240_F2
     misc_feature    1407
                     /label=Linker
     CDS             complement(1413..2675)
                     /codon_start=1
                     /label=GCaMP8m
                     /translation="HHHHHHTRRKKTFKEVATAVKIIGRLMGLKINVYIKADKQKNGIK
                     ANFHIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSVESKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEG
                     DATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQ
                     ERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNLPDQLTEEQ
                     IAEYKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGDGTIDFPE
                     FLTMMARKMKYRDTEEEIREAFGVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIR
                     EADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1413..1439
                     /label=842_R
     primer_bind     1413..1437
                     /label=240_R1
     primer_bind     1969..1988
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2228..2249
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2355..2376)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2654..2693)
                     /label=240_F1
     primer_bind     complement(2654..2675)
                     /label=243_F2
     CDS             complement(2658..2675)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     2659..2689
                     /label=243_R1
     misc_feature    2677..2684
                     /label=Linker
     CDS             complement(2685..3659)
                     /codon_start=1
                     /label=Chronos
                     /translation="METAATMTHAFISAVPSAEATIRGLLSAAAVVTPAADAHGETSNA
                     TTAGADHGCFPHINHGTELQHKIAVGLQWFTVIVAIVQLIFYGWHSFKATTGWEEVYVC
                     VIELVKCFIELFHDVDSPATVYQTNGGAVAWLRYSEWLLTCPVILIHLSNLTGLHEEYS
                     KRTMTILVTDIGNIVWGITAAFTKGPLKILFFMIGLFYGVTCFFQIAKVYIESYHTLPK
                     GVCRKICKIMAYVFFCSWLMFPVMFIAGHEGLGLITPYTSGIGHLILDLIAKNTWGFLG
                     HHLRVKIHEHILIHGDIRKTTTINVAGENMEIETFVDEEEEGGV"
     misc_feature    2791..2807
                     /label=seq primer 2
     primer_bind     complement(3103..3140)
                     /label=NG_GA_F2
     primer_bind     3104..3144
                     /label=NG_GA_R1
     misc_feature    3240
                     /label=M-E:GAG
     misc_feature    3414..3431
                     /label=seq primer 3
     primer_bind     complement(3646..3682)
                     /label=243_F1
     misc_feature    3673..3704
                     /label=lox2722(1)
     misc_feature    3756..3788
                     /label=loxP(1)
     misc_feature    3802..3807
                     /label=HindIII
     misc_feature    3808..4412
                     /label=WPRE
     primer_bind     complement(3867..3887)
                     /label=WPRE-R
     primer_bind     3867..3887
                     /label=WPRE-F
     misc_feature    4451..4663
                     /label=SV40 polyA
     misc_feature    4734..4864
                     /label=3' ITR
     misc_feature    5046..5363
                     /label=f1 origin
     primer_bind     5311..5329
                     /label=f1 ori
     misc_feature    5627..6486
                     /label=Amp
     primer_bind     5779..5798
                     /label=seq primer 7
     primer_bind     6426..6445
                     /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 ggcgcgcctt agaatctggt ggcttcaggg aagtccgtgg cacagctgat gaagctgtca
     1261 atgctggaca tgctccgcat gtcgatgacg ccctccgtgc gtgcgggcag aggggccacg
     1321 gggctgggca tgctgctcat ctccagctcc tctggaggct tgctctgcgg ggccatctcc
     1381 ttggtgttga ggatgggctg ggactgcaat tgcttcgctg tcatcatttg tacaaactct
     1441 tcgtagttta cctgaccatc cccatcgatg tctgcttccc tgatcatttc atcaacctct
     1501 tcatctgtta acttctctcc aaggtttgtc atcacgtggc gaagctctgc tgcactgatg
     1561 tagccattgc catccttatc aaacacaccg aacgcttctc taatttcttc ttccgtgtcc
     1621 ctgtatttca tttttcttgc catcattgtc aggaactcag ggaagtcgat tgtgccgtca
     1681 ccgtcggcat ctacttcatt gatcatgtcc tgcagctctg cttctgtggg gttctgcccc
     1741 agagaccgca tcaccgtccc cagctccttg gttgttattg tcccatcccc gtccttgtca
     1801 aatagggaga aagcctcttt gtattctgcg atctgctctt cagtcagttg gtccggcagg
     1861 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     1921 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     1981 ccgtcgtcct tgaagaagat ggtgcgctcc tggatgtagc cttcgggcat ggcggacttg
     2041 aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc
     2101 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc
     2161 agggtcagct tgccgtaggt ggcatcgccc tcaccctcgc cggacacgct gaacttgtgg
     2221 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     2281 cccttgctca ccatgctccc tccggtaccg cccttgtaca gctcgtccat gccgagagtg
     2341 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     2401 ttcgaaagtt tggactccac gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     2461 ccgatggggg tgttctgctg gtagtggtag gcgagctgca cgccgccgtc ctcgatgttg
     2521 tggcggatgt ggaagttcgc cttgatgccg ttcttctgct tgtcggcctt gatatagacg
     2581 ttgatcttca ggcccatcag gcggccgatg atcttcacgg cggtggccac ctccttgaag
     2641 gtcttcttgc gacgcgtgtg atgatggtga tgatgggcgg ccgccactcc tccctcctcc
     2701 tcctcgtcga cgaaggtctc gatctccatg ttctcgccgg ccacgttgat ggtggttgtc
     2761 ttccggatgt cgccgtggat caggatgtgc tcgtggatct tcactctcag gtggtggccc
     2821 aggaagcccc aagtgttctt ggcgatcaga tccaggatca ggtggccgat tccgctggtg
     2881 taaggtgtga tcaggcccag tccctcgtgt ccggcgatga acatcacggg gaacatcagc
     2941 caagagcaga agaagacgta ggccatgatc ttgcaaatct tccggcagac gcctttgggc
     3001 agggtgtggt agctctcgat atacaccttg gcgatctgga agaagcaagt cacgccgtag
     3061 aacaggccga tcatgaagaa caggatcttc agggggccct ttgtaaaggc ggctgtgatc
     3121 ccccacacga tgttgccgat gtcggtcacc aggatggtca tggtccgctt gctgtactct
     3181 tcgtgcagtc cggtcaggtt gctcaggtgg atcaggatca cggggcaagt caggagccac
     3241 tcgctgtacc gcagccaagc cacggctcct ccgttggtct ggtacactgt ggctgggctg
     3301 tcgacatcgt ggaacagctc gatgaagcac ttgacgagct cgatcacgca gacgtagacc
     3361 tcctcccagc cggttgtggc cttgaagctg tgccaaccgt agaagatgag ctgcacgata
     3421 gccacgatca cggtgaacca ctggagtccc actgcgatct tgtgctgcag ctcggttccg
     3481 tggttgatgt gggggaagca accgtgatcg gctccggctg ttgtggcgtt agaggtttct
     3541 ccgtgagcgt ctgctgctgg tgtcaccact gctgcggcgc tcagcaggcc tctaattgtg
     3601 gcttcggcgc taggcacggc tgagataaag gcgtgggtca ttgtggcggc tgtttccata
     3661 ctagcgctag cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga
     3721 aattatcact gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata
     3781 cgaagttatg aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     3841 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3901 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3961 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4021 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4081 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4141 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4201 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     4261 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4321 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4381 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gacccgggcg gccgcttcga
     4441 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     4501 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4561 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     4621 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt
     4681 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     4741 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4801 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4861 cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4921 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4981 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     5041 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5101 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5161 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5221 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5281 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5341 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5401 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5461 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     5521 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5581 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5641 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5701 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5761 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5821 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5881 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5941 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6001 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6061 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6121 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6181 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6241 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6301 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6361 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6421 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6481 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6541 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6601 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6661 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6721 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6781 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6841 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6901 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6961 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7021 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7081 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7141 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7201 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7261 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatg
//
