LOCUS       sequence-57044-e        8175 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 8175)
  AUTHORS   Butko MT, Yang J, Geng Y, Kim HJ, Jeon NL, Shu X, Mackey MR, 
            Ellisman MH, Tsien RY, Lin MZ
  TITLE     Fluorescent and photo-oxidizing TimeSTAMP tags track protein fates 
            in light and electron microscopy.
  JOURNAL   Nat Neurosci. 2012 Nov;15(12):1742-51. doi: 10.1038/nn.3246. Epub 
            2012 Oct 28.
  PUBMED    23103964
REFERENCE   2  (bases 1 to 8175)
  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: "10.1038/nn.3246"; 
            journalName: "Nat Neurosci"; date: "2012-11"; volume: "15"; issue: 
            "12"; pages: "1742-51"
COMMENT     Sequence Label: pCS6-mPSD95a-TS2-HA
FEATURES             Location/Qualifiers
     source          1..8175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2891..2914
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3641..3667
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(4576..4600)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4601..4620)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4602..4620)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4631..4648)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4631..4647)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4640..4662)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    5052..5186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5089..5108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5143..5162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5312..5767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5399..5418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5609..5630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(6055..6123,6124..6915)
                     /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     complement(6273..6292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    7017..7050
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      7128..7716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7617..7636
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7870..7887
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agcctaggcc gcggccaggc cccccaacat ggactgtctc tgtatagtga caaccaagaa
      661 ataccgctac caagatgaag acacgccccc tctggaacac agcccggccc acctccccaa
      721 ccaggccaat tctccccctg tgattgtcaa cacggacacc ctagaagccc caggatatgt
      781 gaacggaaca gagggggaga tggagtatga ggagatcaca ttggaaaggg gtaactcagg
      841 tctgggcttc agcatcgcag gtggcaccga caacccacac atcggtgacg acccatccat
      901 ctttatcacc aagatcattc ctggtggggc tgcagcccag gatggccgcc tcagggtcaa
      961 cgacagcatc ctgtttgtca atgaagtgga tgtccgggag gtgacccatt cagctgcagt
     1021 ggaggccctc aaagaggcgg gttccatcgt tcgcctctac gtcatgcgcc ggaaaccccc
     1081 agctgagaag atcatagaga tcaagcttat caaagggcct aaaggacttg gcttcagcat
     1141 cgcagggggc gttgggaacc agcacatccc tggagataat agcatctacg taaccaagat
     1201 catcgaagga ggcgctgccc acaaggaggg caggttgcag atcggagaca agatcctggc
     1261 ggtcaacagt gtggggctag aggatgtcat gcatgaggac gccgtggcag ccctgaagaa
     1321 cacatatgac gttgtgtacc taaaggtggc caagcccagc aatgcctacc tgagtgacag
     1381 ctatgctccc ccagacatca caacctcata ttctcagcac ctggacaatg agatcagtca
     1441 tagcagctac ttgggcaccg actaccccac agccatgacc cccacttccc ctcggcgcta
     1501 ctcccccgtg gccaaggacc tgctagggga ggaagatatt ccccgggaac caaggcggat
     1561 cgtgatccat cggggctcca ccggcctggg cttcaacatt gtgggcggcg aggacggtga
     1621 aggcatcttc atctccttca tccttgctgg gggtccagct gacctcagtg gggagctacg
     1681 gaagggggac cagatcctgt cggtcaatgg tgttgacctc cgcaatgcca gtcatgaaca
     1741 ggctgccatt gccctgaaga acgcgggtca gacggtcacg atcatcgctc agtataaacc
     1801 agaagagtat agccgattcg aggccaagat ccatgatctt cgggaacagc ttatgaatag
     1861 tagcctgggc tcagggactg catctctgcg aagcaacccc aagcggggct tctatatcag
     1921 ggccctgttt gactacgaca agaccaagga ctgcggtttc ttgagccagg ccctgagctt
     1981 ccactttggg gatgtgcttc atgtaattga cgccagcgac gaagagtggt ggcaagcgcg
     2041 gcgggtccac tctgacagtg agaccgatga cattggcttc attcccagca aacggcgggt
     2101 cgagcgacga gagtggtcaa ggttaaaggc caaggactgg ggctccagct ctggatcaca
     2161 gggtcgagaa gactcggttc tgagctatga gacggtgacg cagatggaag tgcactacgc
     2221 tcgccccatc atcatccttg ggcctaccaa agaccgtgcc aacgatgatc ttctctccga
     2281 gttccccgac aagtttggat cctgtgtccc tcatacgaca cgtcctaagc gggaatatga
     2341 gatagacggc cgcgattacc actttgtctc ctcccgggag aaaatggaga aggacattca
     2401 ggcgcacaag ttcattgagg ctggccagta caacagccac ctctacggga ccagcgtcca
     2461 gtctgtgcga gaggtagcag agcaggggaa gcactgcatc cttgatgtct cagccaatgc
     2521 cgtgcggcgg ctgcaggcgg cccacctgca ccctatcgcc atcttcatcc gtccccgctc
     2581 cctggagaat gtgctagaga tcaataagcg gatcacagag gagcaagccc ggaaagcctt
     2641 cgacagggcc acgaagctgg agcaggagtt cacggagtgc ttctcagcca tcgtagaggg
     2701 cgacagcttt gaagagatct atcacaaagt gaaacgtgtc atcgaagacc tctcaggccc
     2761 ctacatctgg gtcccagccc gagagagact ggagcccggg cccatggcca gcatgactgg
     2821 tggacagcaa atggggtcga cggaggacgt ggtgtgctgc cactcaatct acggcaagaa
     2881 gaagggtatg gattacaagg atgacgacga taagggctct tccgggacag gctgcgtggt
     2941 catagtgggc aggatcgtct tgtccggatc cggcactagt gcgcccatca cggcgtacgc
     3001 ccagcagacg agaggcctcc tagggtgtat aatcaccagc ctgactggcc gggacaaaaa
     3061 ccaagtggag ggtgaggtcc agatcgtgtc aactgctacc caaaccttcc tggcaacgtg
     3121 catcaatggg gtatgctggg cagtctacca cggggccgga acgaggacca tcgcatcacc
     3181 caagggtcct gtcatccaga tgtataccaa tgtggaccaa gaccttgtgg gctggcccgc
     3241 tcctcaaggt tcccgctcat tgacaccctg tacctgcggc tcctcggacc tttacctggt
     3301 cacgaggcac gccgatgtca ttcccgtgcg ccggcgaggt gatagcaggg gtagcctgct
     3361 ttcgccccgg cccatttcct acttgaaagg ctcctcgggg ggtccgctgt tgtgccccgc
     3421 gggacacgcc gtgggcctat tcagggccgc ggtgtgcacc cgtggagtgg ctaaagcggt
     3481 ggactttatc cctgtggaga acctagagac aaccatgaga tccccggtgt tcacggacaa
     3541 ctcctctcca ccagcagtca ccctgacgca cccaatcacc aaaatcgata gggaggttct
     3601 ctaccaggag ttcgatgaga tggaagagtg ctctcagcac tatccctacg atgtgcccga
     3661 ttacgctggc gcgccttgat tcctgccctg gcttggcctg gactcaccct gcctccacca
     3721 cctgggccct tggtctggac tgaatagccc aagcccttcg taccctagcc tttcttccca
     3781 cccttcttat ttatttcctt tctgactgga tcgagcccat tggagggagg acactcctct
     3841 ctgcatgtat ccctgcaccc cagatcaggg cccctagacc ccaggaacct ggggtctggg
     3901 ggggagctgg gctcctggtt ccgagccctt gctccttaag actcctaccc ctgcccgccc
     3961 ccatgcacac acagacccac tgggggcctc tgccctcccc attttctccc acacacattc
     4021 cagaagtcag ggcccctctc aaggagtacc cgctgtaggg atgcagggcc acaggcttcc
     4081 gcgctctccc gaggcagggg ctggggtcac ccctgcccca tcataactcc ccacgccgtt
     4141 tgagttctcc tttattttct ccattctttt cttctcaaag gtggtttttg gggggagaag
     4201 caggggggct ctcctgaggg tccccccgtc tttcacacac ctcccacctt ttttctttgc
     4261 cggtttgcat gagtggaagg tctaactgtg gctttttttt ttttctggga ttttatttca
     4321 ttttatttga ttttattttt ttgggaaaag gggagggatg ggtctgggga gtggggaatg
     4381 tgggaggggg gtgggggggc aaagggtcag gggttgggtg tccgggagcc aggggaggac
     4441 aggaaatgct gccgccttct gcaatttatt tatttttttc ttttgagaga gtgaaaggaa
     4501 gagacagaca cttgaaaaaa aaaaaaaaag ggcggccgct ctagagtatc cctcgagggg
     4561 cccaagctta cgcgtaccca gctttcttgt acaaagtggt ccctatagtg agtcgtatta
     4621 taagctaggc actggccgtc gttttacaac gtcgtgactg ggaaaactgc tagcttggga
     4681 tctttgtgaa ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga
     4741 tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctagctgcat
     4801 atgcttgctg cttgagagtt ttgcttactg agtatgattt atgaaaatat tatacacagg
     4861 agctagtgat tctaattgtt tgtgtatttt agattcacag tcccaaggct catttcaggc
     4921 ccctcagtcc tcacagtctg ttcatgatca taatcagcca taccacattt gtagaggttt
     4981 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5041 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5101 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5161 tcaatgtatc ttatcatgtc tggatcgatc ctgcattaat gaatcggcca acgcgcgggg
     5221 agaggcggtt tgcgtattgg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     5281 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     5341 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5401 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5461 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5521 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5581 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5641 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5701 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     5761 acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5821 taaatacatt caaatatgta tccgctcatg ccaggtcttg gactggtgag aacggcttgc
     5881 tcggcagctt cgatgtgtgc tggagggaga ataaaggtct aagatgtgcg atagagggaa
     5941 gtcgcattga attatgtgct gtgtagggat cgctggtatc aaatatgtgt gcccacccct
     6001 ggcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6061 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6121 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6181 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6241 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6301 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6361 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6421 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6481 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6541 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6601 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6661 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6721 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6781 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6841 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6901 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6961 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgccataa
     7021 cttcgtataa tgtatgctat acgaagttat ggcatgacca aaatccctta acgtgagttt
     7081 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7141 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7201 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7261 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7321 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7381 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7441 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7501 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7561 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7621 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7681 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7741 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7801 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7861 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7921 ctccccgcgc gttggccgat tcattaatgc agagcttgca attcgcgcgt ttttcaatat
     7981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8041 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     8101 gaaaccatta ttatcatgac attaacctat aaaaataggc gtagtacgag gccctttcac
     8161 tcattagatg catgt
//
