LOCUS       sequence-428994-        8422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NTHL1-mCherry in mammalian cells from a retroviral 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8422)
  AUTHORS   Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, 
            Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, 
            Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas 
            A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
  TITLE     CDK-independent role of D-type cyclins in regulating DNA mismatch 
            repair.
  JOURNAL   Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010.
  PUBMED    38458201
REFERENCE   2  (bases 1 to 8422)
  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: "Mol Cell. 
            2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010."
COMMENT     Sequence Label: NTHL1-mCherry
FEATURES             Location/Qualifiers
     source          1..8422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1642..1945
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1946..2149
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3079..3789
                     /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(3224..3242)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3492..3511
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        3817..4316
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3852..3875)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4249..4268
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4337..4936
                     /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(4337..4356)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4833..4853
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5121..5553
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5656..5678)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5814..5834)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5819..6148
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5999..6134
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6170..6189
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6270..6287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6441..7029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6521..6540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7200..7991,7992..8060))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7823..7842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8061..8165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8233..8251
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8351..8422,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc atagttatta atagtaatca attacggggt cattagttca
     1621 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     1681 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     1741 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     1801 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     1861 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     1921 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     1981 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2041 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2101 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     2161 ccgtcagatc cgctagccgc caccatgtgt agtccgcagg agtccgggcc tctccggaga
     2221 agagaggctg cagcagaagc gaggaaaagc cacagccccg tgaagcgtcc gcggaaagca
     2281 cagagactgc gtgtggccta tgagggctcg gacagtgaga aaggtgaggg ggctgagccc
     2341 ctcaaggtgc cagtctggga gccccaggac tggcagcaac agctggtcaa catccgtgcc
     2401 atgaggaaca aaaaggatgc acctgtggac catctgggga ctgagcactg ctatgactcc
     2461 agtgcccccc caaaggtacg caggtaccag gtgctgctgt cactgatgct ctccagccaa
     2521 accaaagacc aggtgacggc gggcgccatg cagcgactgc gggcgcgggg cctgacggtg
     2581 gacagcatcc tgcagacaga tgatgccacg ctgggcaagc tcatctaccc cgtcggtttc
     2641 tggaggagca aggtgaaata catcaagcag accagcgcca tcctgcagca gcactacggt
     2701 ggggacatcc cagcctctgt ggccgagctg gtggcgctgc cgggtgttgg gcccaagatg
     2761 gcacacctgg ctatggctgt ggcctggggc actgtgtcag gcattgcagt ggacacgcat
     2821 gtgcacagaa tcgccaacag gctgaggtgg accaagaagg caaccaagtc cccagaggag
     2881 acccgcgccg ccctggagga gtggctgcct agggagctgt ggcacgagat caatggactc
     2941 ttggtgggct tcggccagca gacctgtctg cctgtgcacc ctcgctgcca cgcctgcctc
     3001 aaccaagccc tctgcccggc cgcccagggt ctcggcggag gatcgtcgac ggtaccgcgg
     3061 gcccgggatc caccggtcat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     3121 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     3181 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     3241 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     3301 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     3361 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     3421 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     3481 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     3541 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     3601 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     3661 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     3721 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     3781 tacaagtgag cggccgcgac tctagataat tctaccgggt aggggaggcg cttttcccaa
     3841 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc
     3901 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg
     3961 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc
     4021 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg
     4081 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt
     4141 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg
     4201 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc
     4261 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct
     4321 gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     4381 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4441 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4501 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4561 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     4621 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     4681 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     4741 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     4801 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4861 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4921 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     4981 accccatgca tcggcgtctc gatcgagata tcagtggtcc aggctctagt tttgactcaa
     5041 caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt
     5101 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctactagct
     5161 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga
     5221 tcaaggtcag gaacagatgg aacagggtcg atcgacccta gagaaccatc agatgtttcc
     5281 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     5341 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     5401 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     5461 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     5521 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
     5581 cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggctgcctcg cgcgtttcgg
     5641 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5701 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5761 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tagatccggc tgtggaatgt
     5821 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5881 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5941 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6001 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6061 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6121 cttttttgga ggcctaggct tttgcaaaaa gcttactggc ttaactatgc ggcatcagag
     6181 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     6241 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6841 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7501 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7801 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8161 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     8221 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt agcttggcca
     8281 ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta
     8341 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     8401 gttcatagcc catatatgga gt
//
