LOCUS       sequence-191142-        6481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CMYA5 (aa 3859 – 4069) with a N-terminal FLAG tag in pDEST26
            mammalian expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6481)
  TITLE     Shiang Lab CMYA5 project plasmids (will be published)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6481)
  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            "Unpublished"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: CMYA5tnC/pDEST26-N-FLAG
FEATURES             Location/Qualifiers
     source          1..6481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..47)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(201..789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(281..300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(960..1751,1752..1820))
                     /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     1583..1602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1821..1925)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2049..2071)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2171..2190
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    2231..2279
                     /note="synthetic polyadenylation signal"
     CDS             complement(2343..3137)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2454..2473)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3064..3083
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3188..3545)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3202..3337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3354..3545
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3502..3522
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3662..4117)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3799..3820)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4011..4030
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4243..4377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4267..4286)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4321..4340
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4767..4789
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4781..4798
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4782..4798
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4809..4828
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4809..4827
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4860..4884
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5537..5561)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5571..5594)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5695..5719)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(5716..5919)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5745..5765)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5920..6223
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6333..6437
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
       61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
      961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1261 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     1921 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     1981 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     2041 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2101 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     2161 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatacgcg gatccttatc
     2221 gctatcgatt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     2281 cggccatcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     2341 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     2401 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     2461 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     2521 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     2581 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     2641 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     2701 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     2761 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     2821 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     2881 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     2941 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     3001 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     3061 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     3121 ccatcttgtt caatcatggt ggctctagcc ttaagttcga gactgttgtg tcagaagaat
     3181 caagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     3241 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     3301 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     3361 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     3421 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     3481 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
     3541 ctcagaaggt acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
     3601 agatccgcgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3661 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3721 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3781 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3841 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3901 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3961 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4021 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4081 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     4141 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc aatacgcaaa
     4201 ccgcctctcc ccgcgcgttg gccgattcat taatgcagga tcgatccaga catgataaga
     4261 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     4321 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     4381 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     4441 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcatgaaca gactgtgagg
     4501 actgaggggc ctgaaatgag ccttgggact gtgaatctaa aatacacaaa caattagaat
     4561 cactagctcc tgtgtataat attttcataa atcatactca gtaagcaaaa ctctcaagca
     4621 gcaagcatat gcagctagtt taacacatta tacacttaaa aattttatat ttaccttaga
     4681 gctttaaatc tctgtaggta gtttgtccaa ttatgtcaca ccacagaagt aaggttcctt
     4741 cacaaagatc ccaagctagc agttttccca gtcacgacgt tgtaaaacga cggccagtgc
     4801 ctagcttata atacgactca ctatagggag agagctatga cgtcgcatgc acgcgatcaa
     4861 ccactttgta caagaaagct gggtctagat aattccttgt gccttacaga atccgggggc
     4921 tctatcccca ggtgcaaagt acattttcca ggtttctcca gggcaaaggc agggtggaca
     4981 ccctcattaa acctgtgcct gatgatgaag agcaactgct cgctctctgc gttgatgaag
     5041 ataagtctct ggttgttgta gtccagcagg atgcccactc tggctggacg ctcagtcaca
     5101 tgaacatcac tcacaatacc actgtagaaa aatgtgtatc tctgtggctc agagcagtgc
     5161 atgtaccatg aggtctcccc ttgtcctagg gcacctgcct gcacagccga gctggagcag
     5221 atgccgagtc gatatgctgg gcagtctgtg actgtggttt cccagtaatg ctggccacag
     5281 gaaggcagct cctcacccag cactgacggg atttccgtca gccgtctcct ctcaccaaag
     5341 ctgatcactg tcccacttga ggaaatgtgt agagcaggat gagctgtttc tctcaacaag
     5401 agaaatctgg ttcctctggt ggagatgaga gcagcttcac tctgttcact tgtcccaaat
     5461 gcattgatgg ccctcacata gaaaaagtag ttatcattgg gttggaggtt aactttcagc
     5521 tggagtccca tgaaagagcc tgcttttttg tacaaacttg ttgatctaga cttgtcgtca
     5581 tcgtctttgt agtcgtagta cgccatggtc cgcggcctag gctagagtcc ggaggctgga
     5641 tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg
     5701 acggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     5761 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     5821 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     5881 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     5941 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     6001 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     6061 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     6121 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     6181 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgacatgca tctaatgcgt
     6241 gaaagggcct cgtaatacgc ctatttttat aggttaatgt catgataata atggtttctt
     6301 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6361 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6421 attgaaaaag cgcgaattgc aagctctgca ttaatgaatc ggccaacgcg cggggagagg
     6481 c
//
