LOCUS       sequence-206723-        7415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7415)
  AUTHORS   Gross GG, Junge JA, Mora RJ, Kwon HB, Olson CA, Takahashi TT, Liman 
            ER, Ellis-Davies GC, McGee AW, Sabatini BL, Roberts RW, Arnold DB
  TITLE     Recombinant probes for visualizing endogenous synaptic proteins in 
            living neurons.
  JOURNAL   Neuron. 2013 Jun 19;78(6):971-85. doi: 10.1016/j.neuron.2013.04.017.
  PUBMED    23791193
REFERENCE   2  (bases 1 to 7415)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2013.04"; journalName: "Neuron"; date: 
            "2013-06-19- 19"; volume: "78"; issue: "6"; pages: "971-85"
COMMENT     Sequence Label: pCAG_GPHN.FingR-mKate2-IL2RGTC
FEATURES             Location/Qualifiers
     source          1..7415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..679
                     /label=chicken beta-actin promoter
     intron          680..1688
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1612..1634
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             2036..2059
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2078..2770
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     CDS             3170..3295
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the rat zinc finger protein Kid-1 
                     (Witzgall et al., 1994)"
                     /label=KRAB-A
                     /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM"
     intron          3338..3471
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3454..3473
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3480..4068
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3533..3553)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3951..3962)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4095..4112)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4101..4212
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4439..4458)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4504..4559
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4558..4577)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4918..4934)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4931..4953)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4942..4958
                     /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(join(4966..4972,4973..4990,4991..4996))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5011..5032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5091..5286
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5137..5272
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5292..5426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5329..5348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5383..5402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5494..5511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5665..6253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5745..5764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6424..7215,7216..7284))
                     /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"
     primer_bind     7047..7066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7285..7389)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agtcttccac agagtactag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggactat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat gggtcgaggt gagccccacg
      421 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
      481 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggcgcgcgc caggcggggc
      541 ggggcggggg cgagggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
      601 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
      661 gcgaagcgcg cggcgggcgg gagtcgctgc gttgccttcg ccccgtgccc cgctccgcgc
      721 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      781 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc tcgtttcttt
      841 tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc cctttgtgcg ggggggagcg
      901 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc ccgcgctgcc
      961 cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgcg tgtgcgcgag
     1021 gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg ctgcgagggg aacaaaggct
     1081 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcggcg gtcgggctgt
     1141 aacccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg
     1201 ctccgtgcgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg
     1261 gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc
     1321 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat
     1381 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg gcggagccga aatctgggag
     1441 gcgccgccgc accccctcta gcgggcgcgg gcgaagcggt gcggcgccgg caggaaggaa
     1501 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctcca tctccagcct
     1561 cggggctgcc gcagggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
     1621 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
     1681 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1741 tcggtaccgc gggcccggga ttacaatgct cgaagtcaag gaagcatcac caaccagcat
     1801 ccagatcagc tggggcaagt acaaggtcat ggttcgctac taccgcatca cctacggtga
     1861 aactggtggc aatagccctg tccaggaatt caccgtgcct ggcagcaagt ccactgctac
     1921 catcagcagc ctgaaacctg gtgtcgacta taccatcacg gtgtacgccg tcacgatcga
     1981 ccactggaac taccaggacc cgatcccgat ctccatcaac taccgcaccg gatccgatta
     2041 caaggatgac gacgataagg gtagcggctc cagatctgtg agcgagctga ttaaggagaa
     2101 catgcacatg aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc
     2161 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg cggtcgaggg
     2221 cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcaaaac
     2281 cttcatcaac cacacccagg gcatccccga cttctttaag cagtccttcc ccgagggctt
     2341 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     2401 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc
     2461 caacggccct gtgatgcaga agaaaacact cggctgggag gcctccaccg agaccctgta
     2521 ccccgctgac ggcggcctgg aaggcagagc cgacatggcc ctgaagctcg tgggcggggg
     2581 ccacctgatc tgcaacttga agaccacata cagatccaag aaacccgcta agaacctcaa
     2641 gatgcccggc gtctactatg tggacagaag actggaaaga atcaaggagg ccgacaaaga
     2701 gacctacgtc gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact
     2761 ggggcacaga tctagatttc agtgccagat ttgcatgcgc aactttagcc gcaaaagcac
     2821 cctgaccgat catattcgca cccataccgg cgaaaaaccg tttgcgtgcg atatttgcgg
     2881 ccgcaaattt gcggcgcgca gcacccgcac cacccatacc aaaattcata ccggcagcca
     2941 gaaaccgttt cagtgccgca tttgcatgcg caactttagc cgcagcgata gcctgagcaa
     3001 acatattcgc acccataccg gcgaaaaacc gtttgcgtgc gatatttgcg gccgcaaatt
     3061 tgcgcagcgc agcaacctga aagtgcatac caaaattcat ctgcgcggca gccagctgat
     3121 cgatggtgtc gacgctcctg aacaacgtga aggtgcttct caagtttctg tttctgttac
     3181 ttttgaagat gttgctgttc tttttactcg tgatgaatgg aaaaaacttg atctttctca
     3241 acgttctctt tatcgtgaag ttatgcttga aaattattct aatcttgctt ctatggctta
     3301 gcgtacaagt aaagcggccc cggactcgag gccgcaggta agtatcaagg ttacaagaca
     3361 ggtttaagga gaccacatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     3421 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtcgaca
     3481 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3541 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3601 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3661 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3721 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3781 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3841 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     3901 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3961 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4021 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag
     4081 ctcggtacga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4141 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4201 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4261 ggacagcaag ggggaggatt gggaagacaa tagcaggcca gcttttgttc cctttagggt
     4321 taattgcgcg cttggcgcta atcatggtca tagctgtttc cactcctcag gtgcaggctg
     4381 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     4441 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     4501 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     4561 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     4621 gtttggcaac atatgccata tgctggctgc catgaacaaa ggtggctata aagaggtcat
     4681 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     4741 ggttagattt ttttttatat tttgtttgtg ttattttttt ctttaacatc cctaaaattt
     4801 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     4861 gtccctcttc tcttatgaag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     4921 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4981 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5041 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     5101 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5161 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5221 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5281 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5341 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5401 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     5461 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5521 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5581 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5641 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5701 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5761 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5821 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     5881 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5941 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6001 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6061 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6121 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6181 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6241 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6301 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6361 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6421 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6481 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6541 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6601 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6661 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6721 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6781 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6841 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6901 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6961 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7021 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7081 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7141 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7201 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7261 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7321 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7381 ggttccgcgc acatttcccc gaaaagtgcc acctg
//
