LOCUS       sequence-264342-        5400 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Homology arms and mEGFP-linker sequence for N-terminus tagging of 
            human SON.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5400)
  TITLE     Allen Institute for Cell Sciences Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5400)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-82:SON-mEGFP
FEATURES             Location/Qualifiers
     source          1..5400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(61..649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(141..160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(820..1611,1612..1680))
                     /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     1443..1462
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1681..1785)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1853..1871
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1909..1931)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2031..2050
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2244..2266
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2258..2275
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2259..2275
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(3293..3295,3296..3298,3299..4009)
                     /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(3338..3359)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3599..3618)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3946..3967
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5059..5075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5059..5075)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5072..5094)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5083..5099
                     /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(5107..5113,5114..5131,5132..5137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5152..5173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5290..5307)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
       61 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      121 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      181 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      241 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      301 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      361 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      421 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      481 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      541 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      601 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      661 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      721 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      781 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      841 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      901 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      961 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1021 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1081 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1141 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1201 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1261 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1321 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1381 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1441 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1501 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1561 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1621 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1681 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1741 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1801 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1861 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1921 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1981 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     2041 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2101 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     2161 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     2221 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     2281 gacgcgtatt gggatcccgc tcgccggaag tcggcccacc gggaccggcg cgggaccagc
     2341 gcggcaccgc ccctcactgc cccgccctac gacggcttcc ctgccccatt ggtccgcgcg
     2401 gccctgggtc tccgcggcga gaatctagtc tgattccctc ccgccctggg cgctgcctga
     2461 ctgctcccca gccgagacac ccaagcctta gcacgtttat aatgtgacag tggtacgggg
     2521 tgccacgcag tgtttccagg agcgccgggg gggagcgcca aaatgctaaa cccattctta
     2581 gccaattgtg cgccgctcac gagtcagagc tgagcagcta ccgcggaaaa atctcgtgac
     2641 acgcctgact cctgatgaga ctaaaatgca ccgggctttg aatgccggtc gcttaggaca
     2701 tccggcctat tgctaaaaaa aaaaatgagg ctggttatag aacgcactgg ggaagacaca
     2761 gggcaacagc gagttcctcc tctcacatgc cagtgggggc gggggctgaa aaattctctc
     2821 ccggatgcat atccctgtcg aaactgaact tggcgccacc tgagagcgcc tgaaagccgt
     2881 gtgttaaaca ccgggttgcc cttcttgctc ttcttcctct tcgcctttaa tctcaggcac
     2941 ttagcaagcc tgttaaggct cccaaacact gctggggact gacaacttat gcggacacgg
     3001 tcgcctcaag agagacggct cctgtaatgg cgcctgcgtc agacactaac atggcggaca
     3061 tagtcgtgcg cggcgcaaac gtcagcacct ctaccccagc cgctcagaaa cagcagagga
     3121 gagcagggcc gtgctcactg attggtccct cggtggagcg ggccggaccg ggcattccct
     3181 ttggagccaa tcaggaaggc gagagaccga attggccgct gcgcctcctc ccgaggcatg
     3241 ctgggagcct ggaggactag cgaggaggag ttgagagaac ggagcggacg ccatggtgag
     3301 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3361 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3421 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3481 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3541 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3601 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3661 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3721 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     3781 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     3841 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3901 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3961 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaaga gcggcctgag
     4021 aagcgcgaca aacatcgagc agatctttag gtctttcgtc gtcagtaaat tccgggaaat
     4081 tcaacaggag ctttccaggt aaacgcctcc cagattcttc tgctcccaac ggaccgttag
     4141 gccctcacct agccttcttt ggcacctttc ttcggagacg cagtcgttcc ccggctcagg
     4201 ccccgctagg gccccgcctg ggcctgggat ccattttccg ggcccccccc aaccgccccc
     4261 ccagccgctc ccttcctttt ccctcgagga acttcgcctt ttccgaggta acccaagact
     4321 ccctgtttca gaaccgagtt aagatgttca cccttctccc ctgtttgggt cccccctgcc
     4381 gtttagctac tcgtaggccc ggttgtccgc cagggacggg cctagttcct tcctcagctc
     4441 ctcctccgcc gccgcgtatc ccctccccct ctccgcctcc gcggattttg ccggccggtg
     4501 cctatgaccg gaaagggtct ctaacggccg tttcttggtt aatcttaaat cctctttatt
     4561 attgcgagct tcagaccgct tgggggtctg gatctattaa taagattatt cggggcctag
     4621 tgtcctgtag tatctgtcag ccgttctgat agttaaatgt ttcaattaaa acgaactcaa
     4681 actgatgcac ttgctagttt gttaaaatac tcgagataaa gatggaaggg gactttgaaa
     4741 tggttgcgat taattacagc aagtgaaagt ggactttgtg tgtaggtcta ttattttctt
     4801 gctacccctg ccccagttga cacgctattt ttgtcattgc tggtagactt tacataattt
     4861 gcgcttggct tcacaaaatt tgaaggcgaa gagattgtta aagaaatggt tgtttactta
     4921 ggaaaatagt gtgtcatgtt cagcggttgt ctctcgatat ttgcttttgt ataagatact
     4981 catttcttta aagtacagaa gtaggaaaga gatcttgaaa cgtaatccca atggcgcgcc
     5041 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5101 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5161 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5221 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5281 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
//
