LOCUS       sequence-190024-        6481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CMYA5 (aa 3859 – 4069) Leu allele (4063) 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  JOURNAL   Unpublished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            "Unpublished"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: CMYA5tnT/pDEST26-N-FLAG
FEATURES             Location/Qualifiers
     source          1..6481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      86..541
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(173..192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     383..404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        658..1015
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        658..849
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(681..701)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      866..1001
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1066..1860
                     /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(1120..1139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1730..1749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1924..1972
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2013..2032)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2132..2154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2278..2382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2383..2451,2452..3243)
                     /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     complement(2601..2620)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3414..4002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3903..3922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4156..4173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(4247..4351)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4461..4764
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4765..4968
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4919..4939
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4965..4989
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5090..5113
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    5123..5147
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5800..5824)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5856..5875)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5857..5875)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5886..5903)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5886..5902)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5895..5917)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    6307..6441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6344..6363)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6398..6417
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ggtttgcgta ttggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
       61 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
      121 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      181 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      241 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      301 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
      361 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
      421 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
      481 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
      541 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
      601 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
      661 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
      721 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
      781 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      841 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      901 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
      961 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     1021 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     1081 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     1141 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     1201 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     1261 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     1321 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     1381 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     1441 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     1501 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     1561 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     1621 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     1681 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     1741 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     1801 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     1861 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     1921 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     1981 gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2041 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2101 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2161 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2221 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2281 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2341 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2401 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2461 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2521 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2581 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2641 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2701 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2761 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2821 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2881 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2941 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3001 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3061 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3121 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3181 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3241 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3301 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3361 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3421 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3481 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3541 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3601 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3661 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3721 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3781 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3841 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3901 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3961 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4021 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4081 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4141 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     4201 ccgcctctcc ccgcgcgttg gccgattcat taatgcagag cttgcaattc gcgctttttc
     4261 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4321 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4381 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatt acgaggccct
     4441 ttcacgcatt agatgcatgt cgttacataa cttacggtaa atggcccgcc tggctgaccg
     4501 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     4561 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     4621 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     4681 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     4741 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     4801 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     4861 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     4921 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     4981 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     5041 cgatccagcc tccggactct agcctaggcc gcggaccatg gcgtactacg actacaaaga
     5101 cgatgacgac aagtctagat caacaagttt gtacaaaaaa gcaggctctt tcatgggact
     5161 ccagctgaaa gttaacctcc aacccaatga taactacttt ttctatgtga gggccatcaa
     5221 tgcatttggg acaagtgaac agagtgaagc tgctctcatc tccaccagag gaaccagatt
     5281 tctcttgttg agagaaacag ctcatcctgc tctacacatt tcctcaagtg ggacagtgat
     5341 cagctttggt gagaggagac ggctgacgga aatcccgtca gtgctgggtg aggagctgcc
     5401 ttcctgtggc cagcattact gggaaaccac agtcacagac tgcccagcat atcgactcgg
     5461 catctgctcc agctcggctg tgcaggcagg tgccctagga caaggggaga cctcatggta
     5521 catgcactgc tctgagccac agagatacac atttttctac agtggtattg tgagtgatgt
     5581 tcatgtgact gagcgtccag ccagagtggg catcctgctg gactacaaca accagagact
     5641 tatcttcatc aacgcagaga gcgagcagtt gctcttcatc atcaggcaca ggtttaatga
     5701 gggtgtccac cctgcctttg ccctggagaa acctggaaaa tgtactttgc acctggggat
     5761 agagcccctg gattctgtaa ggcacaagga attatctaga cccagctttc ttgtacaaag
     5821 tggttgatcg cgtgcatgcg acgtcatagc tctctcccta tagtgagtcg tattataagc
     5881 taggcactgg ccgtcgtttt acaacgtcgt gactgggaaa actgctagct tgggatcttt
     5941 gtgaaggaac cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa
     6001 agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactagc tgcatatgct
     6061 tgctgcttga gagttttgct tactgagtat gatttatgaa aatattatac acaggagcta
     6121 gtgattctaa ttgtttgtgt attttagatt cacagtccca aggctcattt caggcccctc
     6181 agtcctcaca gtctgttcat gatcataatc agccatacca catttgtaga ggttttactt
     6241 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     6301 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6361 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6421 gtatcttatc atgtctggat cgatcctgca ttaatgaatc ggccaacgcg cggggagagg
     6481 c
//
