LOCUS       sequence-35983-e        4758 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 4758)
  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 4758)
  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: pNLS-LSSmKate2-N1
FEATURES             Location/Qualifiers
     source          1..4758
                     /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"
     polyA_signal    1546..1667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1583..1602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1637..1656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1674..2129)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1811..1832)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2023..2042
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2156..2260
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2262..2619
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2262..2453
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2285..2305)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2470..2605
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2654..3448
                     /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             2654..3448
                     /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(2708..2727)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3318..3337
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3636..3655)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3680..3727
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4056..4644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4545..4564
                     /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 atctcgagct caagcttcga attcgccacc atgtcgagga ggcggcgcag
      661 ctacgagaac gatggtggac aacctcacaa aaggaggaag acgtctccac cggtcgccac
      721 catgagcgag ctgattaagg agaacatgca catgaagctg tacatggaag gcaccgtgaa
      781 caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac
      841 catgagaatc aaggtggtcg agggcggccc tctacccttc gccttcgaca tcttggctac
      901 cagcttcatg tacggcagct acaccttcat caaccacacc cagggcatcc ccgacttctt
      961 taagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg
     1021 cgtgctgacc gctacccagg acaccagcct ccaggacggt tgcctcatct acaacgtcaa
     1081 gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa cactcggctg
     1141 ggaggccggc accgagatgc tgtaccccgc tgacggcggc ctggaaggca gatctgacga
     1201 cgccctgaag ctcgtgggcg ggggccacct gatctgcaac ttgaagagca catacagatc
     1261 caagaaaccc gctaagaatc tcaaggtgcc cggcgtctac tatgtggacc gaagactgga
     1321 aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg ctgtggccag
     1381 atactgcgac ctccctagca aactggggca caagcttaat taaagcggcc gcgactctag
     1441 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1501 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1561 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1621 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     1681 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1741 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     1801 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1861 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1921 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1981 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2041 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2101 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2161 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2221 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     2281 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2341 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2401 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2461 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2521 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2581 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     2641 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2701 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2761 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2821 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2881 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2941 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3001 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3061 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3121 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3181 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3241 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3301 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3361 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3421 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3481 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3541 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3601 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     3661 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     3721 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3781 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3841 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     3901 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3961 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4021 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4081 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4141 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4201 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4261 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4321 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4381 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4441 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4501 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4561 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4621 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4681 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4741 cgtattaccg ccatgcat
//
