LOCUS       sequence-35998-e        4741 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 4741)
  AUTHORS   Piatkevich KD, Hulit J, Subach OM, Wu B, Abdulla A, Segall JE, 
            Verkhusha VV
  TITLE     Monomeric red fluorescent proteins with a large Stokes shift.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Mar 23;107(12):5369-74. Epub 2010 Mar
            8.
  PUBMED    20212155
REFERENCE   2  (bases 1 to 4741)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2010-03-23"; volume: "107(12)"; pages: 
            "5369-74. Epub 2010 Mar 8"
COMMENT     Sequence Label: pMito-LSSmKate2-N1
FEATURES             Location/Qualifiers
     source          1..4741
                     /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             597..671
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     polyA_signal    1529..1650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1566..1585)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1620..1639
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1657..2112)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1794..1815)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2006..2025
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2139..2243
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2245..2602
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2245..2436
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2268..2288)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2453..2588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2637..3431
                     /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             2637..3431
                     /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(2691..2710)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3301..3320
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3619..3638)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3663..3710
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4039..4627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4528..4547
                     /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 gctagcatgt
      601 ccgtcctgac gccgctgctg ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc
      661 cgcgcgccaa gatccattcg ttgggggatc caccggtcgc caccatgagc gagctgatta
      721 aggagaacat gcacatgaag ctgtacatgg aaggcaccgt gaacaaccac cacttcaagt
      781 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtgg
      841 tcgagggcgg ccctctaccc ttcgccttcg acatcttggc taccagcttc atgtacggca
      901 gctacacctt catcaaccac acccagggca tccccgactt ctttaagcag tccttccctg
      961 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
     1021 aggacaccag cctccaggac ggttgcctca tctacaacgt caagatcaga ggggtgaact
     1081 tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc ggcaccgaga
     1141 tgctgtaccc cgctgacggc ggcctggaag gcagatctga cgacgccctg aagctcgtgg
     1201 gcgggggcca cctgatctgc aacttgaaga gcacatacag atccaagaaa cccgctaaga
     1261 atctcaaggt gcccggcgtc tactatgtgg accgaagact ggaaagaatc aaggaggccg
     1321 acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta
     1381 gcaaactggg gcacaagctt aattaaagcg gccgcgactc tagatcataa tcagccatac
     1441 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     1501 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1561 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1621 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     1681 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1741 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1801 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1861 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1921 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1981 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2041 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2101 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2161 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2221 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     2281 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2341 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2401 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2461 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2521 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2581 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     2641 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2701 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2761 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2821 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2881 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2941 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3001 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3061 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3121 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     3181 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3241 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3301 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3361 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3421 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3481 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3541 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3601 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3661 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     3721 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3781 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3841 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     3901 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3961 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4021 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4081 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4141 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     4201 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4261 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4321 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4381 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4441 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4501 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4561 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4621 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4681 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     4741 t
//
