LOCUS       sequence-241928-        6373 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 6373)
  AUTHORS   Yamada KM
  TITLE     Yamada Lab Plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6373)
  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: 
            "unpublished"
COMMENT     Sequence Label: pmCherry Paxillin
FEATURES             Location/Qualifiers
     source          1..6373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1327
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(765..783)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1033..1052
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3168..3289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3205..3224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3259..3278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3296..3751)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3433..3454)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3645..3664
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3778..3882
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3884..4241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3884..4075
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3907..3927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4092..4227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4276..5070
                     /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             4276..5070
                     /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(4330..4349)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4940..4959
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5258..5277)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5302..5349
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5678..6266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6167..6186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
      661 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
      721 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
      781 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
      841 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
      901 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
      961 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     1021 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     1081 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     1141 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     1201 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     1261 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     1321 gtacaagtcc ggactcagat ctcgagctgc ggccgccgga tccaagctta tggacgacct
     1381 cgacgccctg ctggcggact tggagtctac cacctcccac atctccaaac ggcctgtgtt
     1441 cttgtcggag gagaccccct actcataccc aactggaaac cacacatacc aggagattgc
     1501 cgtgccaccc cccgtccccc cacccccgtc cagcgaggcc ctcaatggca caatccttga
     1561 ccccttagac cagtggcagc ccagcggctc ccgattcatc caccagcagc ctcagtcctc
     1621 atcacctgtg tacggctcca gtgccaaaac ttccagtgtc tccaaccctc aggacagtgt
     1681 tggctctccg tgctcccgag tgggtgagga ggagcacgtc tacagcttcc ccaacaagca
     1741 gaaatcagct gagccttcac ccaccgtaat gagcacgtcc ctgggcagca acctttctga
     1801 actcgaccgc ctgctgctgg aactgaacgc tgtacagcat aacccgccag gcttccctgc
     1861 agatgaggcc aactcaagcc ccccgcttcc tggggccctg agccccctct atggtgtccc
     1921 agagactaac agccccttgg gaggcaaagc tgggcccctg acgaaagaga agcctaagcg
     1981 gaatgggggc cggggcctgg aggacgtgcg gcccagtgtg gagagtctct tggatgaact
     2041 ggagagctcc gtgcccagcc ccgtccctgc catcactgtg aaccagggcg agatgagcag
     2101 cccgcagcgc gtcacctcca cccaacagca gacacgcatc tcggcctcct ctgccaccag
     2161 ggagctggac gagctgatgg cttcgctgtc ggatttcaag ttcatggccc aggggaagac
     2221 agggagcagc tcaccccctg gggggccccc gaagcccggg agccagctgg acagcatgct
     2281 ggggagcctg cagtctgacc tgaacaagct gggggtcgcc acagtcgcca aaggagtctg
     2341 cggggcctgc aagaagccca tcgccgggca ggttgtgacc gccatgggga agacgtggca
     2401 ccccgagcac ttcgtctgca cccactgcca ggaggagatc ggatcccgga acttcttcga
     2461 gcgggatgga cagccctact gtgaaaagga ctaccacaac ctcttctccc cgcgctgcta
     2521 ctactgcaac ggccccatcc tggataaagt ggtgacagcc cttgaccgga cgtggcaccc
     2581 tgaacacttc ttctgtgcac agtgtggagc cttctttggt cccgaagggt tccacgagaa
     2641 ggacggcaag gcctactgtc gcaaggacta cttcgacatg ttcgcaccca agtgtggcgg
     2701 ctgcgcccgg gccatcctgg agaactatat ctcagccctc aacacgctgt ggcatcctga
     2761 gtgctttgtg tgccgggaat gcttcacgcc attcgtgaac ggcagcttct tcgagcacga
     2821 cgggcagccc tactgtgagg tgcactacca cgagcggcgc ggctcgctgt gttctggctg
     2881 ccagaagccc atcaccggcc gctgcatcac cgccatggcc aagaagttcc accccgagca
     2941 cttcgtctgt gccttctgcc tcaagcagct caacaagggc accttcaagg agcagaacga
     3001 caagccttac tgtcagaact gcttcctcaa gctcttctgc taggctcgag tctagataac
     3061 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3121 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3181 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3241 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     3301 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3361 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3421 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3481 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3541 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3601 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3661 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3721 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3781 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3841 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3901 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3961 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4081 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4141 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4201 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4261 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4321 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4381 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4441 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4501 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4561 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4621 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4681 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4741 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4801 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4861 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4921 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4981 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5041 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5101 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5161 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5221 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5281 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5341 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5401 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5461 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5521 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5641 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5701 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5761 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5821 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6301 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6361 accgtattac cgc
//
