LOCUS       sequence-482790-        8028 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8028)
  AUTHORS   Scaffidi P, Misteli T
  TITLE     Lamin A-dependent misregulation of adult stem cells associated with 
            accelerated ageing.
  JOURNAL   Nat Cell Biol. 2008 Mar 2. ():.
  PUBMED    18311132
REFERENCE   2  (bases 1 to 8028)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2008 Mar 2. ():."
COMMENT     Sequence Label: pBABE-puro-GFP-progerin
FEATURES             Location/Qualifiers
     source          1..8028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      85..220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             244..843
                     /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(244..263)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     740..760
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1064..1533
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1663..1680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1834..2422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1914..1933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2582..3373,3374..3442))
                     /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     3205..3224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3443..3547)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3578..4047
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4112..4311
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4512..4928
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4858..4880
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4896..4912
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      4967..4976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4973..4975,4976..4978,4979..5689)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4973..4975,4976..4978,4979..5689)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5018..5039)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5279..5298)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5626..5647
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7928..7948)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(7933..8028,1..234)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
       61 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
      121 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
      181 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
      241 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
      301 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
      361 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
      421 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
      481 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
      541 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
      601 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
      661 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
      721 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
      781 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
      841 tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg
      901 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
      961 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     1021 agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
     1081 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
     1141 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     1201 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
     1261 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     1321 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     1381 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
     1441 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     1501 agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg
     1561 gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg
     1621 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1681 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1741 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1801 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1861 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1921 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1981 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2041 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2101 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2161 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2221 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2281 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2341 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2401 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2461 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2521 ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2881 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3181 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3541 ttccgcgcac atttccccga aaagtgccac ctgcagcctg aatatgggcc aaacaggata
     3601 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     3661 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     3721 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     3781 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     3841 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     3901 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     3961 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     4021 tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg
     4081 cccagggacc accgacccac caccgggagg caagctggcc agcaacttat ctgtgtctgt
     4141 ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact
     4201 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
     4261 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
     4321 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     4381 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     4441 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     4501 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     4561 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     4621 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     4681 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     4741 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     4801 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     4861 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     4921 taggcgccgg ccggatccca gtgtggtggt acgtacgcta ccggtcgcca ccatggtgag
     4981 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     5041 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     5101 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     5161 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     5221 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     5281 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5341 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5401 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     5461 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     5521 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5581 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5641 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag
     5701 atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcagcg ctgccaacct
     5761 gccggccatg gagaccccgt cccagcggcg cgccacccgc agcggggcgc aggccagctc
     5821 cactccgctg tcgcccaccc gcatcacccg gctgcaggag aaggaggacc tgcaggagct
     5881 caatgatcgc ttggcggtct acatcgaccg tgtgcgctcg ctggaaacgg agaacgcagg
     5941 gctgcgcctt cgcatcaccg agtctgaaga ggtggtcagc cgcgaggtgt ccggcatcaa
     6001 ggccgcctac gaggccgagc tcggggatgc ccgcaagacc cttgactcag tagccaagga
     6061 gcgcgcccgc ctgcagctgg agctgagcaa agtgcgtgag gagtttaagg agctgaaagc
     6121 gcgcaatacc aagaaggagg gtgacctgat agctgctcag gctcggctga aggacctgga
     6181 ggctctgctg aactccaagg aggccgcact gagcactgct ctcagtgaga agcgcacgct
     6241 ggagggcgag ctgcatgatc tgcggggcca ggtggccaag cttgaggcag ccctaggtga
     6301 ggccaagaag caacttcagg atgagatgct gcggcgggtg gatgctgaga acaggctgca
     6361 gaccatgaag gaggaactgg acttccagaa gaacatctac agtgaggagc tgcgtgagac
     6421 caagcgccgt catgagaccc gactggtgga gattgacaat gggaagcagc gtgagtttga
     6481 gagccggctg gcggatgcgc tgcaggaact gcgggcccag catgaggacc aggtggagca
     6541 gtataagaag gagctggaga agacttattc tgccaagctg gacaatgcca ggcagtctgc
     6601 tgagaggaac agcaacctgg tgggggctgc ccacgaggag ctgcagcagt cgcgcatccg
     6661 catcgacagc ctctctgccc agctcagcca gctccagaag cagctggcag ccaaggaggc
     6721 gaagcttcga gacctggagg actcactggc ccgtgagcgg gacaccagcc ggcggctgct
     6781 ggcggaaaag gagcgggaga tggccgagat gcgggcaagg atgcagcagc agctggacga
     6841 gtaccaggag cttctggaca tcaagctggc cctggacatg gagatccacg cctaccgcaa
     6901 gctcttggag ggcgaggagg agaggctacg cctgtccccc agccctacct cgcagcgcag
     6961 ccgtggccgt gcttcctctc actcatccca gacacagggt gggggcagcg tcaccaaaaa
     7021 gcgcaaactg gagtccactg agagccgcag cagcttctca cagcacgcac gcactagcgg
     7081 gcgcgtggcc gtggaggagg tggatgagga gggcaagttt gtccggctgc gcaacaagtc
     7141 caatgaggac cagtccatgg gcaattggca gatcaagcgc cagaatggag atgatccctt
     7201 gctgacttac cggttcccac caaagttcac cctgaaggct gggcaggtgg tgacgatctg
     7261 ggctgcagga gctggggcca cccacagccc ccctaccgac ctggtgtgga aggcacagaa
     7321 cacctggggc tgcgggaaca gcctgcgtac ggctctcatc aactccactg gggaagaagt
     7381 ggccatgcgc aagctggtgc gctcagtgac tgtggttgag gacgacgagg atgaggatgg
     7441 agatgacctg ctccatcacc accacggctc ccactgcagc agctcggggg accccgctga
     7501 gtacaacctg cgctcgcgca ccgtgctgtg cgggacctgc gggcagcctg ccgacaaggc
     7561 atctgccagc ggctcaggag cccagagccc ccagaactgc agcatcatgt aatctgggac
     7621 ctgccaggca ggggtggggg tggaggcttc ctgcgtcctc ctcacctcat gcccaccccc
     7681 tgccctgcac gtcatgggag ggggcttgaa gccaaagaaa aataaccctt tggttttttt
     7741 cttctgtatt tttttttcta agagaagtta ttttctacag tggttttata ctgaaggaaa
     7801 aacacaagca aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     7861 aaaaaaaaaa agcgtctaga aaccctttag atctgctgca gcgcgccagc acagtggtcg
     7921 accctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     7981 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaag
//
