LOCUS       sequence-124444-        5741 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 5741)
  AUTHORS   Jones SA, Shim SH, He J, Zhuang X
  TITLE     Fast, three-dimensional super-resolution imaging of live cells.
  JOURNAL   Nat Methods. 2011 Jun;8(6):499-508. Epub 2011 May 8.
  PUBMED    21552254
REFERENCE   2  (bases 1 to 5741)
  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"; journalName: "Nat 
            Methods."; date: "2011-06"; volume: "8(6)"; pages: "499-508. Epub 
            2011 May 8"
COMMENT     Sequence Label: pSNAP-CLC-SNAP
FEATURES             Location/Qualifiers
     source          1..5741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             620..1165
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for mammalian expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     regulatory      1190..1199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1859..2404
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for mammalian expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    2529..2650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2566..2585)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2620..2639
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2657..3112)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2794..2815)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3006..3025
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3139..3243
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3245..3602
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3245..3436
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3268..3288)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3453..3588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3637..4431
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3637..4431
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3691..3710)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4301..4320
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4619..4638)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4663..4710
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5039..5627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5528..5547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagga tggacaaaga ctgcgaaatg aagcgcacca ccctggatag
      661 ccctctgggc aagctggaac tgtctgggtg cgaacagggc ctgcacgaga tcaagctgct
      721 gggcaaagga acatctgccg ccgacgccgt ggaagtgcct gccccagccg ccgtgctggg
      781 cggaccagag ccactgatgc aggccaccgc ctggctcaac gcctactttc accagcctga
      841 ggccatcgag gagttccctg tgccagccct gcaccaccca gtgttccagc aggagagctt
      901 tacccgccag gtgctgtgga aactgctgaa agtggtgaag ttcggagagg tcatcagcta
      961 ccagcagctg gccgccctgg ccggcaatcc cgccgccacc gccgccgtga aaaccgccct
     1021 gagcggaaat cccgtgccca ttctgatccc ctgccaccgg gtggtgtcta gctctggcgc
     1081 cgtggggggc tacgagggcg ggctcgccgt gaaagagtgg ctgctggccc acgagggcca
     1141 cagactgggc aagcctgggc tgggtatgaa ttctgcagtc gacggtaccg ccaccatggc
     1201 cgagttggat ccatttggcg atcccgccgg tgcgcccggt ggccccgctc taggaaacgg
     1261 ggtggcgggc gccggcgaag aagacccggc cgcggccttc ttggcgcagc aagagagtga
     1321 gattgcgggc atcgagaacg atgaggcctt cgccatcctg gacggcggcg cccccgggcc
     1381 gcagccacac ggcgagccgc cgggaggtcc ggatgctgtt gacggagtga tgaatggtga
     1441 atactaccag gagagcaatg gtccaacaga cagttacgca gccatttcag aagtggatcg
     1501 attgcagtca gagcctgaaa gtatccgtaa gtggagagaa gagcagacag agcgcctgga
     1561 agccctcgat gccaattctc ggaagcaaga agcggagtgg aaagaaaagg cgataaagga
     1621 gctggaagag tggtatgcga ggcaggacga gcagctacag aagacaaagg caaacaacag
     1681 ggcggcagaa gaagcctttg taaacgacat tgacgagtca tccccaggca ctgagtggga
     1741 gcgggtggcc cgcctgtgtg actttaaccc caagtccagc aaacaggcca aagatgtctc
     1801 tcgcatgcgc tcagtcctca tctccctgaa gcaggccccc ctggtgcacc caccggtgat
     1861 ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca agctggaact
     1921 gtctgggtgc gaacagggcc tgcacgagat caagctgctg ggcaaaggaa catctgccgc
     1981 cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc cactgatgca
     2041 ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg agttccctgt
     2101 gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg tgctgtggaa
     2161 actgctgaaa gtggtgaagt tcggagaggt catcagctac cagcagctgg ccgccctggc
     2221 cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc ccgtgcccat
     2281 tctgatcccc tgccaccggg tggtgtctag ctctggcgcc gtggggggct acgagggcgg
     2341 gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca agcctgggct
     2401 gggttgagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     2461 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2521 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2581 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2641 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2701 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2761 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2821 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2881 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2941 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3001 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3061 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3121 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3181 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3241 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3301 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3361 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3421 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3481 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3541 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3601 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3661 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3721 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3781 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3841 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3901 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3961 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4021 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4081 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4141 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4201 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4261 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4321 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4381 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4441 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4501 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4561 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4621 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4681 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4741 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4801 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4861 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     4921 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4981 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5161 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5701 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
