LOCUS       sequence-418219-        5955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of LynG probe - LynAnchor-eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5955)
  AUTHORS   Tashev SA, Euchner J, Yserentant K, Hanselmann S, Hild F, 
            Chmielewicz W, Hummert J, Schworer F, Tsopoulidis N, Germer S, 
            Sassmannshausen Z, Fackler OT, Klingmuller U, Herten DP
  TITLE     ProDOL: a general method to determine the degree of labeling for 
            staining optimization and molecular counting.
  JOURNAL   Nat Methods. 2024 Sep;21(9):1708-1715. doi: 
            10.1038/s41592-024-02376-6. Epub 2024 Aug 8.
  PUBMED    39117875
REFERENCE   2  (bases 1 to 5955)
  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"; doi: 
            "10.1038/s41592-024-02376-6"; journalName: "Nat Methods"; date: 
            "2024-09"; volume: "21"; issue: "9"; pages: "1708-1715"
COMMENT     Sequence Label: pBABE-LynG
FEATURES             Location/Qualifiers
     source          1..5955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(496..1287,1288..1356))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1119..1138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1357..1461)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1492..1961
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2026..2225
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     primer_bind     2772..2794
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     CDS             join(2990..2992,2993..2995,2996..3706)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3035..3056)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3296..3315)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3643..3664
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3717..3734
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3769..3789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3774..4103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3954..4089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4113..4712
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4113..4132)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4609..4629
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4933..5402
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5532..5549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5703..5955,1..336))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5783..5802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
       61 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      181 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      421 aaaaggatct tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta
      481 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
      541 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      601 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
      661 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
      721 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
      781 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
      841 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
      901 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      961 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1021 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1081 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1141 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1201 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1261 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1321 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1381 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1441 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag cctgaatatg
     1501 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     1561 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     1621 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     1681 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     1741 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     1801 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     1861 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     1921 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga
     1981 tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggcaagct ggccagcaac
     2041 ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta
     2101 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
     2161 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
     2221 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
     2281 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
     2341 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt
     2401 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     2461 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     2521 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     2581 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     2641 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     2701 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc
     2761 cccgtctctc ccccttgaac ctcctcgttc gaccccccct cgagctgcag agcttgctga
     2821 cccagctgtg gactagggga aaggggaaag aggaaaggga aggagaggtg ggggggacga
     2881 aacaggggcg ctggagccct gagcttcctg aagctagggc tgcatcgaat tcatgggatg
     2941 tatcaagagt aagcgtaagg ataatctcaa tgacgacgag ctcgagacca tggtgagcaa
     3001 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3061 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3121 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3181 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     3241 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3301 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3361 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3421 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3481 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3541 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     3601 ccagtccaaa ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3661 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggat caattgcacc
     3721 atcaccatca ccactaatta attaagccag cacagtggtc gaccctgtgg aatgtgtgtc
     3781 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3841 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3901 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3961 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4021 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4081 ttggaggcct aggcttttgc aaaaagctta ccatgaccga gtacaagccc acggtgcgcc
     4141 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     4201 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     4261 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     4321 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     4381 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     4441 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     4501 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     4561 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     4621 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     4681 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     4741 ccgaaaggag cgcacgaccc catgcatcga taaaataaaa gattttattt agtctccaga
     4801 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     4861 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     4921 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     4981 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     5041 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     5101 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     5161 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     5221 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     5281 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     5341 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     5401 cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg
     5461 ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5521 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5581 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5641 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5701 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5761 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5821 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5881 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5941 tccaagctgg gctgt
//
