LOCUS       sequence-400611-        5885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5885)
  TITLE     Eukaryotic expression plasmid with a puromycin selection marker
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5885)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1-Pur-mEGFP
FEATURES             Location/Qualifiers
     source          1..5885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    130..251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(167..186)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     221..240
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(300..316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(300..316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(313..335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    324..340
                     /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(348..354,355..372,373..378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    393..414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(531..548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(702..1290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(782..801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1461..2252,2253..2321))
                     /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     2084..2103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2322..2426)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2501..2520)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2692..3071
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3072..3275
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3226..3246
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3320..3339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3320..3338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3384..3386,3387..3389,3390..4097)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLKE
                     RVTAAGITLGMDELYK"
     primer_bind     complement(3426..3447)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3687..3706)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(4175..4192)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4181..4405
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4451..4879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4538..4557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4748..4769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4888..4908)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4893..5222
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5073..5208
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5135..5154
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5278..5877
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
ORIGIN
        1 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
       61 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
      121 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
      181 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
      241 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
      301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
      421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
      481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
      541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1141 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1441 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1501 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1561 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1621 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1681 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1741 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1801 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1861 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1921 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1981 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2041 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2101 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2161 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2221 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2281 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2341 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2401 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     2461 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     2521 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     2581 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     2641 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     2701 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     2761 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     2821 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     2881 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     2941 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     3001 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     3061 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     3121 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     3181 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     3241 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
     3301 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt
     3361 taaacgggcc ctctagactc gagatggtga gcaagggcga gctgttcacc ggggtggtgc
     3421 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3481 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3541 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3601 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3661 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3721 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3781 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3841 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3901 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3961 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     4021 agaagcgcga tcacatggtc ctgaaggagc gcgtgaccgc cgccgggatc actctcggca
     4081 tggacgagct gtacaaggat atctgcagaa ttccaccaca ctggactagt ggatccgagc
     4141 tcggtaccaa gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag
     4201 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     4261 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     4321 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     4381 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     4441 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4501 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4561 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4621 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4681 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4741 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4801 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4861 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4921 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4981 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5041 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5101 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5161 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5221 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg cctgaggatg
     5281 accgagtaca agcctaccgt gcgcctggcc actcgcgatg atgtgccccg cgccgtccgc
     5341 actctggccg ccgctttcgc cgactacccc gctacccggc acaccgtgga ccccgaccgg
     5401 cacatcgagc gtgtgacaga gttgcaggag ctgttcctga cccgcgtcgg gctggacatc
     5461 ggcaaggtgt gggtagccga cgacggcgcg gccgtggccg tgtggactac ccccgagagc
     5521 gttgaggccg gcgccgtgtt cgccgagatc ggcccccgaa tggccgagct gagcggcagc
     5581 cgcctggccg cccagcagca aatggagggc ctgcttgccc cccatcgtcc caaggagcct
     5641 gcctggtttc tggccactgt aggagtgagc cccgaccacc agggcaaggg cttgggcagc
     5701 gccgtcgtgt tgcccggcgt agaggccgcc gaacgcgccg gtgtgcccgc ctttctcgaa
     5761 acaagcgcac caagaaacct tccattctac gagcgcctgg gcttcaccgt gaccgccgat
     5821 gtcgaggtgc ccgagggacc taggacctgg tgtatgacac gaaaacctgg cgcctaaggc
     5881 gcgcc
//
