LOCUS       sequence-36001-e        6039 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 6039)
  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 6039)
  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: pLSSmKate1-Tubulin-C1
FEATURES             Location/Qualifiers
     source          1..6039
                     /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"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2827..2948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2864..2883)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2918..2937
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2955..3410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3092..3113)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3304..3323
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3437..3541
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3543..3900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3543..3734
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3566..3586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3751..3886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3935..4729
                     /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             3935..4729
                     /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(3989..4008)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4599..4618
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4917..4936)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4961..5008
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5337..5925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5826..5845
                     /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 gctagcggtg
      601 gccaccatgg tgtctaaggg cgaagagctg attaaggaga acatgcacat gaagctgtac
      661 atggaaggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc
      721 tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct acccttcgcc
      781 ttcgacatct tggctaccag cttcatgtac ggcagctaca ccttcatcaa ccacacccag
      841 ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga gagagtcacc
      901 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggttgc
      961 ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag
     1021 aagaaaacac tcggctggga ggccgggacc gagatgctgt accccgctga cggcggcctg
     1081 gaaggcagaa gcgacgaggc cctgaagctc gtgggcgggg gccacctgat ctgcaacttg
     1141 aagagcacat acagatccaa gaaacccgct aagaatctca aggtgcccgg cgtctactat
     1201 gtggaccgaa gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac
     1261 gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacag aggtggagga
     1321 ggttccggac tcagatctcg agtgcgtgag tgcatctcca tccacgttgg ccaggctggt
     1381 gtccagattg gcaatgcctg ctgggagctc tactgcctgg aacacggcat ccagcccgat
     1441 ggccagatgc caagtgacaa gaccattggg ggaggagatg actccttcaa caccttcttc
     1501 agtgagacgg gcgctggcaa gcacgtgccc cgggctgtgt ttgtagactt ggaacccaca
     1561 gtcattgatg aagttcgcac tggcacctac cgccagctct tccaccctga gcagctcatc
     1621 acaggcaagg aagatgctgc caataactat gcccgagggc actacaccat tggcaaggag
     1681 atcattgacc ttgtgttgga ccgaattcgc aagctggctg accagtgcac cggtcttcag
     1741 ggcttcttgg ttttccacag ctttggtggg ggaactggtt ctgggttcac ctccctgctc
     1801 atggaacgtc tctcagttga ttatggcaag aagtccaagc tggagttctc catttaccca
     1861 gcaccccagg tttccacagc tgtagttgag ccctacaact ccatcctcac cacccacacc
     1921 accctggagc actctgattg tgccttcatg gtagacaatg aggccatcta tgacatctgt
     1981 cgtagaaacc tcgatatcga gcgcccaacc tacactaacc ttaaccgcct tattagccag
     2041 attgtgtcct ccatcactgc ttccctgaga tttgatggag ccctgaatgt tgacctgaca
     2101 gaattccaga ccaacctggt gccctacccc cgcatccact tccctctggc cacatatgcc
     2161 cctgtcatct ctgctgagaa agcctaccat gaacagcttt ctgtagcaga gatcaccaat
     2221 gcttgctttg agccagccaa ccagatggtg aaatgtgacc ctcgccatgg taaatacatg
     2281 gcttgctgcc tgttgtaccg tggtgacgtg gttcccaaag atgtcaatgc tgccattgcc
     2341 accatcaaaa ccaagcgcag catccagttt gtggattggt gccccactgg cttcaaggtt
     2401 ggcatcaact accagcctcc cactgtggtg cctggtggag acctggccaa ggtacagaga
     2461 gctgtgtgca tgctgagcaa caccacagcc attgctgagg cctgggctcg cctggaccac
     2521 aagtttgacc tgatgtatgc caagcgtgcc tttgttcact ggtacgtggg tgaggggatg
     2581 gaggaaggcg agttttcaga ggcccgtgaa gatatggctg cccttgagaa ggattatgag
     2641 gaggttggtg tggattctgt tgaaggagag ggtgaggaag aaggagagga atactaagga
     2701 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     2761 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2821 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2881 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2941 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3001 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3061 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3121 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3181 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3241 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3301 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3361 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3421 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3481 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3541 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3661 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3721 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3781 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3841 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3901 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3961 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4021 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4081 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4141 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4201 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4261 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4321 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4381 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4441 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4501 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4561 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4621 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4681 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4741 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4801 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4861 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4921 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4981 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5041 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5101 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5161 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5221 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5281 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5341 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5401 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5461 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5521 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5581 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5641 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5701 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5761 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5821 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5881 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5941 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6001 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
