LOCUS       sequence-301933-        6316 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 6316)
  TITLE     Drapkin Lab plasmids
REFERENCE   2  (bases 1 to 6316)
  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: piRES-puro-ELF3
FEATURES             Location/Qualifiers
     source          1..6316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          2161..2390
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2373..2392
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2462..3036,3037..3039,3040..3048)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2629..2646)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2856..2875
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3068..3667
                     /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(3068..3087)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3564..3584
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3803..3820)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3809..4033
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4080..4099
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4159..4175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4159..4175)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4172..4194)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4183..4199
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4207..4213,4214..4231,4232..4237))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4252..4273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4390..4407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4561..5149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4641..4660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5320..6111,6112..6180))
                     /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     5943..5962
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6181..6285)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtcgtacgt
      961 ctccggattc gaattcatgg ctgcaacctg tgagattagc aacattttta gcaactactt
     1021 cagtgcgatg tacagctcgg aggactccac cctggcctct gttccccctg ctgccacctt
     1081 tggggccgat gacttggtac tgaccctgag caacccccag atgtcattgg agggtacaga
     1141 gaaggccagc tggttggggg aacagcccca gttctggtcg aagacgcagg ttctggactg
     1201 gatcagctac caagtggaga agaacaagta cgacgcaagc gccattgact tctcacgatg
     1261 tgacatggat ggcgccaccc tctgcaattg tgcccttgag gagctgcgtc tggtctttgg
     1321 gcctctgggg gaccaactcc atgcccagct gcgagacctc acttccagct cttctgatga
     1381 gctcagttgg atcattgagc tgctggagaa ggatggcatg gccttccagg aggccctaga
     1441 cccagggccc tttgaccagg gcagcccctt tgcccaggag ctgctggacg acggtcagca
     1501 agccagcccc taccaccccg gcagctgtgg cgcaggagcc ccctcccctg gcagctctga
     1561 cgtctccacc gcagggactg gtgcttctcg gagctcccac tcctcagact ccggtggaag
     1621 tgacgtggac ctggatccca ctgatggcaa gctcttcccc agcgatggtt ttcgtgactg
     1681 caagaagggg gatcccaagc acgggaagcg gaaacgaggc cggccccgaa agctgagcaa
     1741 agagtactgg gactgtctcg agggcaagaa gagcaagcac gcgcccagag gcacccacct
     1801 gtgggagttc atccgggaca tcctcatcca cccggagctc aacgagggcc tcatgaagtg
     1861 ggagaatcgg catgaaggcg tcttcaagtt cctgcgctcc gaggctgtgg cccaactatg
     1921 gggccaaaag aaaaagaaca gcaacatgac ctacgagaag ctgagccggg ccatgaggta
     1981 ctactacaaa cgggagatcc tggaacgggt ggatggccgg cgactcgtct acaagtttgg
     2041 caaaaactca agcggctgga aggaggaaga ggttctccag agtcggaact gagcggccgc
     2101 atagataact gatccagtgt gctggaatta attcgctgtc tgcgagggcc agctgttggg
     2161 gtgagtactc cctctcaaaa gcgggcatga cttctgcgct aagattgtca gtttccaaaa
     2221 acgaggagga tttgatattc acctggcccg cggtgatgcc tttgagggtg gccgcgtcca
     2281 tctggtcaga aaagacaatc tttttgttgt caagcttgag gtgtggcagg cttgagatct
     2341 ggccatacac ttgagtgaca atgacatcca ctttgccttt ctctccacag gtgtccactc
     2401 ccaggtccaa ctgcaggtcg agcatgcatc tagggcggcc aattccgccc ctctcccccc
     2461 ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     2521 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     2581 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     2641 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     2701 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     2761 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     2821 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     2881 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     2941 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     3001 gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaaccca caaggagacg
     3061 accttccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg
     3121 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     3181 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     3241 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     3301 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     3361 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     3421 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     3481 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     3541 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     3601 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     3661 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgg
     3721 ctccgaccga agccacccgg ggcggccccg ccgaccccgc acccgccccc gaggcccacc
     3781 gactctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3841 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3901 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3961 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4021 gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgagtgc attctagttg
     4081 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4141 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4201 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4261 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4321 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4381 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4441 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4501 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4561 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4621 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4681 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4741 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4801 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4861 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4921 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4981 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5041 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5101 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5161 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5221 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5281 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5341 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5401 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5461 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5521 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5581 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5641 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5701 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5761 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5821 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5881 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5941 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6001 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6061 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6121 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6181 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6241 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6301 agtgccacct gacgtc
//
