LOCUS       sequence-137772-        7897 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 7897)
  AUTHORS   Yamada KM
  TITLE     Yamada Lab Plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7897)
  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            "unpublished"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 Vinculin
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      562..571
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             568..1275
                     /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(713..731)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     981..1000
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4640..4761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4677..4696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4731..4750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4768..5223)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4905..4926)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5117..5136
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5250..5354
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5356..5713
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5356..5547
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5379..5399)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5564..5699
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5748..6542
                     /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             5748..6542
                     /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(5802..5821)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6412..6431
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6730..6749)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6774..6821
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7150..7738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7639..7658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc
      601 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      661 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      721 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
      781 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
      841 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
      901 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
      961 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1021 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     1081 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     1141 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     1201 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     1261 gacgagctgt acaagtccgg actcagatct cgagctgcgg ccgccggatc caagcttatg
     1321 cccgtcttcc acacgcgcac catcgagagc atcttggagc ccgtggctca gcagatctcc
     1381 cacctggtca tcatgcacga ggagggggag gtagacggca aggccatccc ggacctcacc
     1441 gcccccgtgt cggccgtgca ggccgctgtc agcaacctgg tgcgggttgg aaaagaaact
     1501 gtgcagacaa cagaagacca gatcttgaaa agggatatgc caccagcatt catcaaagta
     1561 gagaatgcct gcaccaagct cgttcgagca gcccagatgc tgcaagcaga tccttattca
     1621 gtaccagctc gtgactacct aattgatgga tcaagaggca tcctttctgg aacatcagac
     1681 ttacttctga catttgatga agcggaggtc cgtaaaatca tccgtgtctg caaaggaata
     1741 ttggaatatc tgactgtggc agaagtagta gagactatgg aggatttggt gacatataca
     1801 aagaatctag ggccaggaat gacaaagatg gcgaaaatga tcgatgagag acaacaggaa
     1861 ttaactcatc aggaacatag ggttatgctg gtgaactcca tgaatactgt gaaggagcta
     1921 ttgcctgtac ttatttcagc tatgaagatc tttgtaacca caaaaaacac taaaagccag
     1981 ggaatagaag aggccttgaa aaatcgcaat ttcacagtag agaaaatgag tgctgagata
     2041 aatgaaataa tccgtgtatt acaactcact tcctgggatg aagatgcctg ggccagcaag
     2101 gacactgaag ccatgaaaag agctttagcc ctaatagatt caaagatgaa ccaggcaaaa
     2161 ggctggctga gagatccaaa cgcacctcca ggggatgctg gtgagcaagc aataaggcag
     2221 atccttgatg aagctggaaa agcaggagaa ttgtgtgcag gcaaagaacg cagggagatt
     2281 ctgggaacat gcaaaacttt gggccaaatg actgatcaac ttgctgatct ccgagctaga
     2341 ggacagggtg ctacaccgat ggcaatgcag aaggcacagc aggtgtcaca aggcctggat
     2401 ttgctcactg caaaagtgga gaatgcagcc cggaaattgg aggccatgac aaactctaag
     2461 caggctattg caaagaagat tgatgctgct cagaactggc ttgcggatcc caacggcgga
     2521 agtgaaggag aagaacacat tcgaggaatt atgtctgaag caaggaaagt tgcagaattg
     2581 tgtgaggagc ctaaagaaag agatgatatc cttcgctcct tgggggaaat ctctgctctg
     2641 acagctaagc tgtcagatct gcgacgacat gggaaaggcg actctcctga ggcccgtgca
     2701 ttggccaagc aaatagctac atcacttcag aacttacagt ccaaaacaaa cagagctgta
     2761 gcaaatacta ggccagttaa agctgctgtc catttggagg gcaagattga gcaagcccag
     2821 aggtggatag ataatcctac agttgatgat cggggagtag gccaggcagc aattcggggt
     2881 ttggttgcag aaggtcgtcg tctagccaat gtcatgatgg gaccttatcg tcaggacctg
     2941 cttgccaaat gtgaccgtgt agaccagctg gctgctcagc ttgctgacct tgcagcaaga
     3001 ggggaaggag agtctcctca ggctagggca attgctgctc agcttcagga ctccctgaag
     3061 gatctcaaag cacggatgca agaagcaatg acccaggagg tgtctgatgt tttcagtgac
     3121 actacaaccc ctattaagtt gttagcagta gcagccactg ctccttctga tactcccaat
     3181 agagaagagg tgtttgaaga aagagcagca aattttgaaa accatgctgc tagactggga
     3241 gcaacagcag aaaaagcagc tgcagttgga actgctaata aaactactgt ggaaggcatt
     3301 caggcaacag tcaaatctgc aagggagctt acaccacagg tagtatcggc tgctcgaatc
     3361 ctcctgagaa atcctggaaa tcaagctgct tatgagcatt ttgagacaat gaaaaaccaa
     3421 tggattgata atgtagaaaa gatgacaggg ctggtggatg aggccatcga taccaagtct
     3481 ctgttggatg catcagaaga ggctattaag aaggatcttg ataaatgtaa agttgcaatg
     3541 gccaatatgc aacctcagat gctggtagct ggagccacca gcattgctag acgagcgaac
     3601 cgcatcctgc ttgtggcaaa acgggaggtt gaaaattcag aagaccctaa attcagggag
     3661 gctgttaaag cagcttctga tgagctgagc aaaaccatat caccgatggt aatggatgct
     3721 aaagctgtag caggaaatat ctctgatcct ggtttgcaga agagtttctt ggattctgga
     3781 tacaggattc tgggagctgt ggccaaagtc agagaagcct ttcagcctca ggagccagac
     3841 tttccccctc ctcctcctga cctagagcat ctccatctga ctgatgagct tgctcctcca
     3901 aaaccaccac ttccagaagg tgaggttccc ccacccagac caccaccccc tgaagaaaaa
     3961 gatgaggagt tcccagagca gaaagcagga gaagctatta atcagcccat gatgatggct
     4021 gctaggcagt tgcatgacga ggctcggaaa tggtctagca agggtaacga catcattgct
     4081 gctgctaaac gaatggcgct gctcatggct gagatgtcac gcctggtgcg aggaggcagc
     4141 ggaaacaagc gtgccctcat ccagtgtgca aaagatattg ctaaggcatc ggatgaagtc
     4201 actcgattgg ccaaagaggt ggccaagcaa tgtactgata aacgcattag aacaaacctc
     4261 ttacaggtct gtgagcgaat cccaaccatc agcacgcagc tcaaaattct ttccacagtg
     4321 aaagctacca tgctgggcag gactaacatc agcgatgaag aatcagaaca ggcaactgag
     4381 atgttggttc ataacgccca gaacctcatg cagtctgtga aggaaactgt gagagaagct
     4441 gaagcagcat ccattaagat aagaacagat gccggattca ctctgcgctg ggtcagaaag
     4501 accccatggt atcagtaaac actctagata actgatcata atcagccata ccacatttgt
     4561 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     4621 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4681 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4741 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     4801 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4861 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4921 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4981 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     5041 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     5101 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5161 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     5221 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5281 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5341 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     5401 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5461 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5521 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     5581 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5641 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5701 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5761 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5821 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5881 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     5941 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     6001 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     6061 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     6121 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     6181 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6241 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     6301 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6361 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6421 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6481 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6541 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6601 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6661 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     6721 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     6781 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     6841 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     6901 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     6961 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     7021 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7081 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7141 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7201 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7261 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7321 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7381 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7441 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7501 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7561 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7621 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7681 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7741 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7801 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     7861 aatagtaatc aattacgggg tcattagttc atagccc
//
