LOCUS       sequence-373227-        5080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of human MYF5 ORF with a N-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5080)
  TITLE     Alexander lab Muscle Factors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5080)
  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            "Unpublished"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: FLAG-hMYF5 (#795)
FEATURES             Location/Qualifiers
     source          1..5080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        554..572
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             616..639
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1443..1459)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1444..1460)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1515..1534)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1516..1534)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1808..1929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1845..1864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1899..1918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1936..2391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2073..2094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2285..2304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2418..2522
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2524..2881
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2524..2715
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2547..2567)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2732..2867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2916..3710
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2916..3710
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2970..2989)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3580..3599
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3898..3917)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3942..3989
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4318..4906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4807..4826
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt
      541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggtg
      601 gcggccgcca ccatggatta caaggatgac gacgataaga gcccgggcgg atccatggac
      661 gtgatggatg gctgccagtt ctcaccttct gagtacttct acgacggctc ctgcataccg
      721 tcccccgagg gtgaatttgg ggacgagttt gtgccgcgag tggctgcctt cggagcgcac
      781 aaagcagagc tgcagggctc agatgaggac gagcacgtgc gagcgcctac cggccaccac
      841 caggctggtc actgcctcat gtgggcctgc aaagcctgca agaggaagtc caccaccatg
      901 gatcggcgga aggcagccac tatgcgcgag cggaggcgcc tgaagaaggt caaccaggct
      961 ttcgaaaccc tcaagaggtg taccacgacc aaccccaacc agaggctgcc caaggtggag
     1021 atcctcagga atgccatccg ctacatcgag agcctgcagg agttgctgag agagcaggtg
     1081 gagaactact atagcctgcc gggacagagc tgctcggagc ccaccagccc cacctccaac
     1141 tgctctgatg gcatgcccga atgtaacagt cctgtctggt ccagaaagag cagtactttt
     1201 gacagcatct actgtcctga tgtatcaaat gtatatgcca cagataaaaa ctccttatcc
     1261 agcttggatt gcttatccaa catagtggac cggatcacct cctcagagca acctgggttg
     1321 cctctccagg atctggcttc tctctctcca gttgccagca ccgattcaca gcctgcaact
     1381 ccaggggctt ctagttccag gcttatctat catgtgctat gagaattcga tatcaagctt
     1441 atcgataccg tcgacctcga gggggggccc ggtaccttaa ttaattaagg taccaggtaa
     1501 gtgtacccaa ttcgccctat agtgagtcgt attacaattc actcgatcgc ccttcccaac
     1561 agttgcgcag cctgaatggc gaatggagat ccaattttta agtgtataat gtgttaaact
     1621 actgattcta attgtttgtg tattttagat tcacagtccc aaggctcatt tcaggcccct
     1681 cagtcctcac agtctgttca tgatcataat cagccatacc acatttgtag aggttttact
     1741 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     1801 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1861 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1921 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     1981 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2041 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2101 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2161 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2221 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2281 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2341 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2401 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2461 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2521 atcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2581 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2641 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2701 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2761 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2821 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2881 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     2941 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3001 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3061 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3121 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3181 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3241 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3301 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3361 aagccggtct tgtcgatcag gatgatctgg acgaagaaca tcaggggctc gcgccagccg
     3421 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3481 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3541 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3601 ctgaagaact tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3661 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3721 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3781 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3841 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     3901 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     3961 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4021 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4081 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4141 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4201 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4261 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4321 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4381 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4441 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4501 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4561 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4621 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4681 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4741 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4801 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4861 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4921 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4981 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     5041 ttacggggtc attagttcat agcccatata tggagttccg
//
