LOCUS       sequence-169265-        6116 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 6116)
  TITLE     Kyriakis lab plasmids
REFERENCE   2  (bases 1 to 6116)
  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"
COMMENT     Sequence Label: pMT3 p38
FEATURES             Location/Qualifiers
     source          1..6116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     152..171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     365..387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     379..396
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     380..396
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(406..735)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      420..555
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     720..740
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          1204..1433
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1416..1435
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1468..1494
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     RBS             2501..2509
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2716..3279
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    3466..3600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3503..3522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3557..3576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4127..4144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4298..4886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4378..4397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5057..5848,5849..5917))
                     /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     5680..5699
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5918..6022)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6090..6108
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
      241 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
      301 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
      361 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctttttgc aaaagcctag
      421 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
      481 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
      541 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
      601 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
      661 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
      721 ccctaactga cacacattcc acaggatccg gtcgcgcgaa tttcgagcgg tgttccgcgg
      781 tcctcctcgt atagaaactc ggaccactct gagacgaagg ctcgcgtcca ggccagcacg
      841 aaggaggcta agtgggaggg gtagcggtcg ttgtccacta gggggtccac tcgctccagg
      901 gtgtgaagac acatgtcgcc ctcttcggca tcaaggaagg tgattggttt ataggtgtag
      961 gccacgtgac cgggtgttcc tgaagggggg ctataaaagg gggtgggggc gcgttcgtcc
     1021 tcactctctt ccgcatcgct gtctgcgagg gccagctgtt gggctcgcgg ttgaggacaa
     1081 actcttcgcg gtctttccag tactcttgga tcggaaaccc gtcggcctcc gaacggtact
     1141 ccgccaccga gggacctgag cgagtccgca tcgaccggat cggaaaacct ctcgactgtt
     1201 ggggtgagta ctccctctca aaagcgggca tgacttctgc gctaagattg tcagtttcca
     1261 aaaacgagga ggatttgata ttcacctggc ccgcggtgat gcctttgagg gtggccgcgt
     1321 ccatctggtc agaaaagaca atctttttgt tgtcaagctt gaggtgtggc aggcttgaga
     1381 tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca caggtgtcca
     1441 ctcccaggtc caactgcagc caccatgtac ccatacgatg ttccagatta cgctggaatt
     1501 cctctagaga tgtctcagga gaggcccacg ttctaccggc aggagctgaa caagacaatc
     1561 tgggaggtgc ccgagcgtta ccagaacctg tctccagtgg gctctggcgc ctatggctct
     1621 gtgtgtgctg cttttgacac aaaaacgggg ttacgtgtgg cagtgaagaa gctctccaga
     1681 ccatttcagt ccatcattca tgcgaaaaga acctacagag aactgcggtt acttaaacat
     1741 atgaaacatg aaaatgtgat tggtctgttg gacgttttta cacctgcaag gtctctggag
     1801 gaattcaatg atgtgtatct ggtgacccat ctcatggggg cagatctgaa caacattgtg
     1861 aaatgtcaga agcttacaga tgaccatgtt cagttcctta tctaccaaat tctccgaggt
     1921 ctaaagtata tacattcagc tgacataatt cacagggacc taaaacctag taatctagct
     1981 gtgaatgaag actgtgagct gaagattctg gattttggac tggctcggca cacagatgat
     2041 gaaatgacag gctacgtggc cactaggtgg tacagggctc ctgagatcat gctgaactgg
     2101 atgcattaca accagacagt tgatatttgg tcagtgggat gcataatggc cgagctgttg
     2161 actggaagaa cattgtttcc tggtacagac catattgatc agttgaagct cattttaaga
     2221 ctcgttggaa ccccaggggc tgagcttttg aagaaaatct cctcagagtc tgcaagaaac
     2281 tatattcagt ctttgactca gatgccgaag atgaactttg cgaatgtatt tattggtgcc
     2341 aatcccctgg ctgtcgactt gctggagaag atgcttgtat tggactcaga taagagaatt
     2401 acagcggccc aagcccttgc acatgcctac tttgctcagt accatgatcc tgatgatgaa
     2461 ccagtggccg atccttatga tcagtccttt gaaagcaggg acctccttat agatgagtgg
     2521 aaaagcctga cctatgatga agtcatcagc tttgtgccac caccccttga ccaagaagag
     2581 atggagtcct gagcacctgg tttgcggccg caattccggg gggggggggg gggacagctc
     2641 agggctgcga tttcgcgcca aacttgacgg caatcctagc gtgaaggctg gtaggatttt
     2701 atccccgctg ccatcatggt tcgaccattg aactgcatcg tcgccgtgtc ccaaaatatg
     2761 gggattggca agaacggaga cctaccctgg cctccgctca ggaacgagtt caagtacttc
     2821 caaagaatga ccacaacctc ttcagtggaa ggtaaacaga atctggtgat tatgggtagg
     2881 aaaacctggt tctccattcc tgagaagaat cgacctttaa aggacagaat taatatagtt
     2941 ctcagtagag aactcaaaga accaccacga ggagctcatt ttcttgccaa aagtttggat
     3001 gatgccttaa gacttattga acaaccggaa ttggcaagta aagtagacat ggtttggata
     3061 gtcggaggca gttctgttta ccaggaagcc atgaatcaac caggccacct cagactcttt
     3121 gtgacaagga tcatgcagga atttgaaagt gacacgtttt tcccagaaat tgatttgggg
     3181 aaatataaac ttctcccaga atacccaggc gtcctctctg aggtccagga ggaaaaaggc
     3241 atcaagtata agtttgaagt ctacgagaag aaagactaac aggaagatgc tttcaagttc
     3301 tctgctcccc tcctaaagct atgcattttt ataagaccat gggacttttg ctggctttag
     3361 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3421 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3481 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3541 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     3601 cccggccaac ggtctggtga cccggctgcg agagctcggt gtacctgaga cgcgagtaag
     3661 cccttgagtc aaagacgtag tcgttgcaag tccgcaccag gtactgatat cccaccaaaa
     3721 agtgcggcgg cggctggcgg tagaggggcc agcgtagggt ggccggggct ccgggggcga
     3781 ggtcttccaa cataaggcga tgatcatcga tgctagaccg tgcaaaagga gagcctgtaa
     3841 gcgggcactc ttccgtggtc tggtggataa attcgcaagg gtatcatggc ggacgaccgg
     3901 ggttcgaacc ccggatccgg ccgtccgccg tgatccatgc ggttaccgcc cgcgtgtcga
     3961 acccaggtgt gcgacgtcag acaacggggg agcgctcctt ttggcttcct tccaggcgcg
     4021 gcggctgctg cgctagcttt tttggcgagc tcgaattaat tctgcattaa tgaatcggcc
     4081 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4141 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4201 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4261 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4321 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4381 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4441 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4501 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4561 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4621 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4681 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4741 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4801 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4861 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4921 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4981 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5041 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5101 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5161 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5221 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5281 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5341 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5401 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5461 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5521 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5581 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5641 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5701 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5761 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5821 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5881 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5941 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6001 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     6061 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgt
//
