LOCUS       sequence-380539-        5394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mEGFP-SRA for N-terminus tagging of human  DCP1A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5394)
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5394)
  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            "Unpublished"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: AICSDP-079:DCP1A-mEGFP
FEATURES             Location/Qualifiers
     source          1..5394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1793..1811
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1849..1871)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1971..1990
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2184..2206
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2198..2215
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2199..2215
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(3233..3235,3236..3238,3239..3949)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3278..3299)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3539..3558)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3886..3907
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4993..5009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4993..5009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5006..5028)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5017..5033
                     /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(5041..5047,5048..5065,5066..5071))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5086..5107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5224..5241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1801 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     1981 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2041 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     2101 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     2161 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     2221 gacgcgtatt gggatttcaa aaagaaaaaa agaaaagggc cgggcgcggt ggctcaagcc
     2281 tgtaatctca gcactttggg aggcagaggc gggtggatcg cctgaggtca ggagttcgag
     2341 actagcctgg ccaacatggc gaaaccccgt ctctactata aacacaaaaa ttatcagggc
     2401 atggtggcgc gcccatgtaa tcccagctac tggggacgct gagacaggag aatcgcttga
     2461 accttggagg cggaggttgc agtgagtcga gatcgcgcca ctgcacttca gcctgggcga
     2521 caagagtgaa actccgtctc aaaaaaaaag aaaaaaaatc cacttgtacc ttctgctaat
     2581 cagtgtatat attcaaggca acttgaatct atgctcctgg ggtgcaatcc tcaagcttgg
     2641 cccaaataaa ctctctgctt gcatttattt tgccccagag tcttcttcct tttaggtcga
     2701 cacatccata tctgagacct agggtaattt tctgggcttc tcgcctctcc ctttccccca
     2761 tgtttttctg tgctctctaa acgggcccgg tagagtcccg catttccaca ttccacagac
     2821 atgttttcct tctgtggccc cgggccctgc cagacagcgc accgcgaggc cggcacttcg
     2881 gcggtctcca acctttgcct ggaaagagct cggcaggcga gctagaggcc agatcccagg
     2941 acccagtcgt tttagctcag ggaaaggaag cgccggagct ccagcctgca agctccacta
     3001 cgcagccttg gacaccgccc acatcgtagg gccgcggacc ctgacaacgc cggaacccgg
     3061 cgcccggagc gtgcgcgcgg cggaccagaa tggctgacgt accgccctgc cgcgaggccc
     3121 gcgtagaggc ggaagttgag gggactgacg cagatggggg aattttgcct cgatggcact
     3181 ttcctgtccg cgactccgcc cccaccagag gggctaggct ctgtgattca agatggtcag
     3241 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3301 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3361 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3421 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3481 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3541 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3601 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3661 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     3721 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     3781 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3841 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3901 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaaga gcagagccga
     3961 ggcgctgagt cgggctgggc aggagatgag cctagcggcc ctgaagcaac acgaccccta
     4021 tatcaccagc atcgcagacc tcacgggcca ggtcgctctg tacaccttct gccccaaggc
     4081 caaccagtgg gtgagtgccg cctggccctg aggacggcca cccggccgct gccgtctctt
     4141 aaaggggccg cgcgggttac tgtggggtgg gggggcacac taagaggcca gggcggtgga
     4201 gacgaggcca gggtctggcc aagagcctag ccagagcctg aggggtgtcg ggtccgcttg
     4261 ggattggggg ataggagtga gagaagtggc tcgagacatc ccaaggaaaa gccagtggag
     4321 gggaggattg gagcaggatc agtcccttgg cgtcgggtct ggggacgtgg gcgacctgcc
     4381 ttctttgcag atacattggg agttataaga ccggtggggt caaaagggtg cctgaaagct
     4441 tgtgtttgag gctttgcgga ttattagata aattatatgt acattatcta taaaataaac
     4501 actcctcctc tccccctggc aaacgtaaat gagagcccct gtcgcagagg tcatttgggt
     4561 ttaaagttta aaatccaatt agtttcgatt cttggaagga gggggtgggt gggtccggct
     4621 ttaatcgaac tacacagagt ttcggaaaag attccaaggc acttaggaaa agggcttccg
     4681 tgttcactgg atccccgaag acctttgtac aaacaagaga gaccctgtaa atgtgttttg
     4741 ttccatctat gatgctggtg atgtcatctc tgaatgctcc cctcggactc agtactttct
     4801 ggctttcgtg cactggggag ggtccgaaaa gtcacattca gtgtgttcac tgattcttgt
     4861 tagcccctgc ttgaagcttt gttagttgtt taaaaagcat aatttcccac tttcttgtaa
     4921 ttgacccttc ctaattaact cattcagcta caggctttat cccaatggcg cgccgagctt
     4981 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5041 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5101 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5161 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5221 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5281 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5341 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtt
//
