LOCUS       CaMK2rep                6661 bp    DNA     circular UNA 16-APR-2025
DEFINITION  Gibson Assembly ligation product.
ACCESSION   urn.local...y7-f8tn41v
VERSION     urn.local...y7-f8tn41v
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     source          join(<1821..6661,1..>1094)
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     enhancer        235..614
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /label="CMV promoter"
     primer_bind     769..789
                     /%GC="66.7"
                     /Hairpin_Tm="52.4"
                     /Self_Dimer_Tm="3.5"
                     /Tm="67.0"
                     /annotation_group="56322239171610007478338-4008-4765-8dcf-
                     82d847cdef96"
                     /created_by="primer3"
                     /Sequence="CGCAAATGGGCGGTAGGCGTG"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="CaMK2rep"
                     /label="CMV fw"
     promoter        863..881
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     source          <954..>1088
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     endo_site       984..989
                     /created_by="User"
                     /label="KpnI"
     CDS             990..>1088
                     /created_by="User"
                     /label="Spot-NES-mCherry"
     endo_site       999..1034
                     /created_by="User"
                     /label="Spot"
     misc_feature    1041..1082
                     /created_by="User"
                     /label="NES"
                     /note="Geneious type: nuclear export signal"
     misc_feature    1081..1834
                     /Source="referenceToDocument: urn:local:.:5h-c68te6h"
                     /label="pSGFP2-C1 PCR Product (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     source          <1089..>1094
                     /organism="pRSET-B"
                     /mol_type="other DNA"
                     /label="source pRSET-B"
     endo_site       1089..1094
                     /created_by="User"
                     /label="EcoRI"
     sig_peptide     <1101..1814
                     /label="SGFP2"
     misc_feature    1813..1102
                     /Source="referenceToDocument: urn:local:.:4x-c68tb7v"
                     /label="pcDNA3.1 reporter PCR Product (concatenated
                     sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    <1821..2228
                     /Original_Bases="CACCACACTGGACTAGTGGATCCGAGCTCGGTACC"
                     /label="CACCACACTGGACTAGTGGATCCGAGCTCGGTACC"
                     /note="Geneious type: Editing History Replacement"
     sequence        1821..>2057
                     /created_by="User"
                     /label="SynP3"
     CDS             <1821..2225
                     /codon_start=1
                     /created_by="User"
                     /modified_by="User"
                     /label="mCherry-SynP3myc"
     phospho_site    1875..1907
                     /created_by="User"
                     /modified_by="User"
                     /label="Syn phospho site 3"
     primer_bind     complement(1875..1907)
                     /%GC="69.7"
                     /Hairpin_Tm="60.6"
                     /Self_Dimer_Tm="26.0"
                     /Tm="79.6"
                     /annotation_group="5172009177143001bfb2e00-1ae1-441e-ac2b-
                     134392a7122e"
                     /created_by="primer3"
                     /Sequence="ACCGGGCCCTGCCTGAGCAGCCTGTCGGATTGG"
                     /Mismatch_Positions="================GC==============="
                     /Mismatches=2
                     /#_Off-target_Sites=0
                     /Off-target_Template="CaMK2AR wt"
                     /label="Rv-SynP3_site1 (S/A)"
     primer_bind     1875..1907
                     /%GC="69.7"
                     /Hairpin_Tm="58.2"
                     /Self_Dimer_Tm="23.8"
                     /Tm="79.6"
                     /annotation_group="51722180588720092721756-2469-4731-b983-
                     72297afb2bbe"
                     /created_by="primer3"
                     /Sequence="CCAATCCGACAGGCTGCTCAGGCAGGGCCCGGT"
                     /Mismatch_Positions="===============GC================"
                     /Mismatches=2
                     /#_Off-target_Sites=0
                     /Off-target_Template="CaMK2AR wt"
                     /label="Fw-SynP3_site1 (S/A)"
     mismatch        1890
                     /Target_base="A"
                     /Oligo_base="G"
                     /Oligo_name="Fw-SynP3 (S/A)"
                     /annotation_group="516714654200800d9d3c7aa-3ad3-4392-b676-
                     788fa043a6d2"
                     /label="A/G"
     mismatch        1890
                     /Target_base="T"
                     /Oligo_base="C"
                     /Oligo_name="Rv-SynP3 (S/A)"
                     /annotation_group="516936462472000e8b3a809-7bc6-44ac-a624-
                     e9136882ad27"
                     /label="T/C"
     mismatch        1890
                     /Target_base="T"
                     /Oligo_base="C"
                     /Oligo_name="Rv-SynP3_site1 (S/A)"
                     /annotation_group="5172009177143001bfb2e00-1ae1-441e-ac2b-
                     134392a7122e"
                     /label="T/C"
     mismatch        1890
                     /Target_base="A"
                     /Oligo_base="G"
                     /Oligo_name="Fw-SynP3_site1 (S/A)"
                     /annotation_group="51722180588720092721756-2469-4731-b983-
                     72297afb2bbe"
                     /label="A/G"
     mismatch        1891
                     /Target_base="G"
                     /Oligo_base="C"
                     /Oligo_name="Fw-SynP3 (S/A)"
                     /annotation_group="516714654200800d9d3c7aa-3ad3-4392-b676-
                     788fa043a6d2"
                     /label="G/C"
     mismatch        1891
                     /Target_base="C"
                     /Oligo_base="G"
                     /Oligo_name="Rv-SynP3 (S/A)"
                     /annotation_group="516936462472000e8b3a809-7bc6-44ac-a624-
                     e9136882ad27"
                     /label="C/G"
     mismatch        1891
                     /Target_base="C"
                     /Oligo_base="G"
                     /Oligo_name="Rv-SynP3_site1 (S/A)"
                     /annotation_group="5172009177143001bfb2e00-1ae1-441e-ac2b-
                     134392a7122e"
                     /label="C/G"
     mismatch        1891
                     /Target_base="G"
                     /Oligo_base="C"
                     /Oligo_name="Fw-SynP3_site1 (S/A)"
                     /annotation_group="51722180588720092721756-2469-4731-b983-
                     72297afb2bbe"
                     /label="G/C"
     phospho_site    1986..2021
                     /created_by="User"
                     /modified_by="User"
                     /label="Syn phospho site 3"
     linker          2058..2069
                     /created_by="User"
                     /modified_by="User"
                     /label="flexible linker"
     endo_site       2058..2063
                     /created_by="User"
                     /label="BamHI"
     tag             2070..2099
                     /created_by="User"
                     /label="myc tag"
     linker          2100..2123
                     /created_by="User"
                     /label="flexible linker"
     tag             2124..2153
                     /created_by="User"
                     /modified_by="User"
                     /label="myc tag"
     linker          2154..2177
                     /created_by="User"
                     /modified_by="User"
                     /label="flexible linker"
     tag             2178..2207
                     /created_by="User"
                     /label="myc tag"
     endo_site       2217..2222
                     /created_by="User"
                     /label="BamHI"
     endo_site       2229..2234
                     /created_by="User"
                     /label="HindIII"
     polyA_signal    2261..2485
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal polyA signal"
     primer_bind     complement(2286..2305)
                     /%GC="65.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="65.2"
                     /annotation_group="563350881158700abea6bae-6d7d-4bdb-b3e0-
                     8b200556c680"
                     /created_by="primer3"
                     /Sequence="CGGGGGAGGGGCAAACAACA"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="CaMK2rep"
                     /label="TGB reverse seq primer"
     primer_bind     complement(2389..2417)
                     /%GC="58.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="70.7"
                     /annotation_group="563351013585100fa5db9a8-de89-41da-925f-
                     fde53b832dca"
                     /created_by="primer3"
                     /Sequence="CCCACCCCACCCCCCAGAATAGAATGACA"
                     /Mismatches=0
                     /#_Off-target_Sites=0
                     /Off-target_Template="CaMK2rep"
                     /label="oligo 3' pcDNA3"
     rep_origin      2531..2959
                     /direction="RIGHT"
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori rep origin"
     promoter        2973..3302
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     rep_origin      3153..3288
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
                     /label="SV40 ori rep origin"
     CDS             3369..4163
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and
                     G418(Geneticin)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
                     /label="aph(3')-II (or nptII) CDS"
     polyA_signal    4337..4458
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal polyA signal"
     rep_origin      complement(4909..5494)
                     /direction="LEFT"
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori rep origin"
     CDS             complement(5665..6525)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="bla CDS"
     promoter        complement(6526..6630)
                     /gene="bla"
                     /note="AmpR promoter"
                     /label="bla promoter"
ORIGIN      
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcaggaatgg
      961 tgagcaaggg cgaggaggac aacggtacca tgggtagccc cgatagggta cgagctgtta
     1021 gccactggag ctcaggctct agtagattgc agttgcctcc attggaaaga ctgaccctcg
     1081 ataagactga attcaccggt gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1141 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1201 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1261 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcgcccgct
     1321 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1381 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1441 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1501 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     1561 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     1621 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1681 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     1741 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1801 acgagctgta caagcctagg gcagcacgac ctccggcgtc ccctagccca caacgccaag
     1861 ccggtccacc tcagccaatc cgacaggcta gtcaggcagg gcccggtcca cctaaagtgt
     1921 caggggcgtc tcccggcggg caacaaagac agggtcctcc acaaaaaccg ccgggcccag
     1981 caggtcctat ccgacaggct tcccaggctg ggcctggccc aagaactggc cctcccacga
     2041 cgcaacagcc cagacccgga tccgggggtg agcaaaaact tatatctgag gaggatcttg
     2101 gaggtggcgg aagtggaggt ggagaacaaa agctaatatc agaggaggat ttgggaggtg
     2161 gcggaagtgg aggtggagag cagaagctga tctctgaaga ggacctcggc ggaggtggat
     2221 cctagtaaaa gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag
     2281 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2341 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2401 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2461 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2521 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2581 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2641 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2701 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2761 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2821 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2881 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2941 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3061 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3121 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3181 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3241 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3301 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3361 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3421 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3481 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3541 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3601 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3661 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3721 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3781 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3841 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3901 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3961 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4021 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4081 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4141 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4201 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4261 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4321 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4381 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4441 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4501 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4561 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4621 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4681 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4741 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4801 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4861 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4921 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4981 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5041 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5101 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5161 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5221 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5281 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5341 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5401 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa
     5461 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5521 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5581 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5641 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5701 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5761 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5821 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5881 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5941 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6001 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6061 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6121 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6181 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6241 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6301 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6361 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6421 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6481 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6541 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6601 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6661 c
//
