LOCUS       Exported                5955 bp DNA     circular SYN 13-JUL-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5955)
  AUTHORS   Chapman Lab
  TITLE     Direct Submission
  JOURNAL   Exported Jul 13, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5955
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        36..1217
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          266..1208
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1151..1174
                     /label=pEF_seq_F
     CDS             1234..2157
                     /codon_start=1
                     /label=Synaptophysin
                     /translation="MDVVNQLVAGGQFRVVKEPLGFVKVLQWVFAIFAFATCGSYTGEL
                     RLSVECANKTESALNIEVEFEYPFRLHQVYFDAPSCVKGGTTKIFLVGDYSSSAEFFVT
                     VAVFAFLYSMGALATYIFLQNKYRENNKGPMMDFLATAVFAFMWLVSSSAWAKGLSDVK
                     MATDPENIIKEMPMCRQTGNTCKELRDPVTSGLNTSVVFGFLNLVLWVGNLWFVFKETG
                     WAAPFMRAPPGAPEKQPAPGDAYGDAGYGQGPGGYGPQDSYGPQGGYQPDYGQPASGGG
                     GYGPQGDYGQQGYGQQGAPTSFSNQMK"
     misc_feature    2158..2199
                     /label=GS(GSS)4 Linker1
     primer_bind     complement(2168..2205)
                     /label=Physin-GSS-Tw4_IF_rev
     primer_bind     complement(2175..2216)
                     /label=Physin-GSS-Tw2B_IF_rev
     primer_bind     complement(2179..2199)
                     /label=physin-GCaMP_IF_rev
     CDS             2200..2895
                     /codon_start=1
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGG"
     primer_bind     complement(2924..2942)
                     /label=pEF Seq R
     polyA_site      3044..3099
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(3458..3474)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3482..3498
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3506..3536)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3551..3572
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3631..3826
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3677..3812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3832..3966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4205..4793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4964..5824)
                     /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"
     promoter        complement(5825..5929)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agatcatcgc gtgaggctcc ggtgcccgtc agtgggcaga
       61 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
      121 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
      181 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
      241 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
      301 ctttacgggt tatggccctt gcgtgccttg aattacttcc acgcccctgg ctgcagtacg
      361 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
      421 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
      481 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
      541 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
      601 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
      661 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
      721 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
      781 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
      841 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
      901 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
      961 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     1021 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     1081 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     1141 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     1201 catttcaggt gtcgtgagga attgcccgcc accatggacg tggtgaatca gctggtggct
     1261 gggggtcagt tccgggtggt caaggagccc cttggcttcg tgaaggtgct gcagtgggtc
     1321 tttgccatct tcgcctttgc tacgtgtggc agctacaccg gggagcttcg gctgagcgtg
     1381 gagtgtgcca acaagacgga gagtgccctc aacatcgaag ttgaattcga gtaccccttc
     1441 aggctgcacc aagtgtactt tgatgcaccc tcctgcgtca aagggggcac taccaagatc
     1501 ttcctggttg gggactactc ctcgtcggct gaattctttg tcaccgtggc tgtgtttgcc
     1561 ttcctctact ccatgggggc cctggccacc tacatcttcc tgcagaacaa gtaccgagag
     1621 aacaacaaag ggcctatgat ggactttctg gctacagccg tgttcgcttt catgtggcta
     1681 gttagttcat cagcctgggc caaaggcctg tccgatgtga agatggccac ggacccagag
     1741 aacattatca aggagatgcc catgtgccgc cagacaggga acacatgcaa ggaactgagg
     1801 gaccctgtga cttcaggact caacacctca gtggtgtttg gcttcctgaa cctggtgctc
     1861 tgggttggca acttatggtt cgtgttcaag gagacaggct gggcagcccc attcatgcgc
     1921 gcacctccag gcgccccgga aaagcaacca gcacctggcg atgcctacgg cgatgcgggc
     1981 tacgggcagg gccccggagg ctatgggccc caggactcct acgggcctca gggtggttat
     2041 caacccgatt acgggcagcc agccagcggt ggcggtggct acgggcctca gggcgactat
     2101 gggcagcaag gctatggcca acagggtgcg cccacctcct tctccaatca gatgaaagga
     2161 tccggaagtt cggggtccag cggttcttca ggcagttcca tggtgtctaa gggcgaagag
     2221 ctgatcaagg aaaatatgcg tatgaaggtg gtcatggaag gttcggtcaa cggccaccaa
     2281 ttcaaatgca caggtgaagg agaaggcaga ccgtacgagg gagtgcaaac catgaggatc
     2341 aaagtcatcg agggaggacc cctgccattt gcctttgaca ttcttgccac gtcgttcatg
     2401 tatggcagcc gtacctttat caagtacccg gccgacatcc ctgatttctt taaacagtcc
     2461 tttcctgagg gttttacttg ggaaagagtt acgagatacg aagatggtgg agtcgtcacc
     2521 gtcacgcagg acaccagcct tgaggatggc gagctcgtct acaacgtcaa ggtcagaggg
     2581 gtaaactttc cctccaatgg tcccgtgatg cagaagaaga ccaagggttg ggagcctaat
     2641 acagagatga tgtatccagc agatggtggt ctgagaggat acactgacat cgcactgaaa
     2701 gttgatggtg gtggccatct gcactgcaac ttcgtgacaa cttacaggtc aaaaaagacc
     2761 gtcgggaaca tcaagatgcc cggtgtccat gccgttgatc accgcctgga aaggatcgag
     2821 gagagtgaca atgaaaccta cgtagtgcaa agagaagtgg cagttgccaa atacagcaac
     2881 cttggtggtg gctagggccg cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     2941 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     3001 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     3061 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     3121 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgccata
     3181 tgctggctgc catgaacaaa ggtggctata aagaggtcat cagtatatga aacagccccc
     3241 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     3301 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     3361 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag
     3421 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3481 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3541 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3601 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     3661 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3721 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3781 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     3841 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3901 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3961 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4021 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4081 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4141 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4201 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4261 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4321 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4381 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     4441 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4501 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4561 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4621 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4681 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4741 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4801 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4861 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4921 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4981 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5041 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5101 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5161 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5221 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5281 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5341 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5401 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5461 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5521 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5581 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5641 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5701 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5761 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5821 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5881 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5941 gaaaagtgcc acctg
//
