LOCUS       sequence-387125-        4160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3xFlag mammalian expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4160)
  TITLE     tag plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4160)
  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            "Unpublished"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: pCS2+N-3xFlag Cloning Vector
FEATURES             Location/Qualifiers
     source          1..4160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             81..146
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    212..346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(236..255)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     290..309
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(459..479)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(459..477)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(498..514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(498..514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(511..533)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    522..538
                     /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(546..552,553..570,571..576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    591..612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(729..746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(900..1488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(980..999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1659..2450,2451..2519))
                     /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     2282..2301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2520..2624)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2650..3105)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2787..2808)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2999..3018
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3176..4160
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agacttgttc
       61 tttttgcagg atctaccatg gactacaaag accatgacgg tgattataaa gatcatgaca
      121 tcgattacaa ggatgacgat gacaagggat cccatcgatt cgaattcaag gcctctcgag
      181 cctctagaac tatagtgagt cgtattacgt agatccagac atgataagat acattgatga
      241 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      301 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      361 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc
      421 ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
      481 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
      541 ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc
      601 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
      661 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
      721 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      781 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      841 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      901 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      961 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1021 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1081 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1141 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1201 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1261 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1321 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1381 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1441 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1501 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1561 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     1621 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     1681 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     1741 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     1801 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1861 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1921 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     1981 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2041 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2101 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2161 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2221 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     2281 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     2341 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     2401 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     2461 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     2521 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     2581 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     2641 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2701 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2761 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2821 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2881 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2941 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3001 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3061 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     3121 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga
     3181 ccgccaattc aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct
     3241 gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc
     3301 cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg acctggcact
     3361 gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc
     3421 tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat
     3481 atgccaggat caatatatag gcaatatcca atatggccct atgccaatat ggctattggc
     3541 caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta
     3601 ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat
     3661 accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg
     3721 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
     3781 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
     3841 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
     3901 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
     3961 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
     4021 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
     4081 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
     4141 gcaatgctcg tttagggaac
//
