LOCUS       sequence-310669-        7390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of MSTN gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7390)
  TITLE     Bi Lab mammalian expression constructs
REFERENCE   2  (bases 1 to 7390)
  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"
COMMENT     Sequence Label: p3x-FLAG-CMV-MSTN
FEATURES             Location/Qualifiers
     source          1..7390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2073..2138
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2149..2771
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2348..2367)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2795..2815)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2813..3110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3164..3958
                     /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(3218..3237)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3828..3847
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4610..4744
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4647..4666)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4701..4720
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4749..4768)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4750..4768)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4782..4798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4782..4798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4795..4817)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4806..4822
                     /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(4830..4836,4837..4854,4855..4860))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4875..4896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5013..5030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5184..5772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5264..5283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5943..6734,6735..6803))
                     /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     6566..6585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6804..6908)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6935..7363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7022..7041)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7232..7253
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaagcttg ccaccatgca aaaactgcaa atctatgttt
      961 atatttacct gtttatgctg attgttgctg gtcccgtgga tctgaatgag aacagcgagc
     1021 aaaaggaaaa tgtggaaaaa gaggggctgt gtaatgcatg tatgtggaga caaaacacta
     1081 aatcttcaag actagaagcc ataaaaattc aaatcctcag taaacttcgc ctggaaacag
     1141 ctcctaacat tagcaaagat gctataagac aacttttgcc caaagctcct ccactccggg
     1201 aactgattga tcagtacgat gtccagagag atgacagcag tgatggctcc ttggaagatg
     1261 atgattatca cgctacgacg gaaacgatca ttaccatgcc tacagagtct gatcttctaa
     1321 tgcaagtgga aggaaaaccc aaatgctgct tctttaaatt tagctctaaa atacaataca
     1381 ataaagtagt aaaggcccaa ctgtggatat atctgagacc cgtcaagact cctacaacag
     1441 tgtttgtgca aatcctgaga ctcatcaaac ccatgaaaga cggtacaagg tatactggaa
     1501 tccgatctct gaaacttgac atgaacccag gcactggtat ttggcagagc attgatgtga
     1561 agacagtgtt gcaaaattgg ctcaaacaac ctgaatccaa cttaggcatt gaaatcaaag
     1621 ctttagatga gaatggtcat gatcttgctg taaccttccc aggaccagga gaagatgggc
     1681 tgaatccctt tttagaagtc aaggtaacag acacaccaaa aagatccagg agagattttg
     1741 gactcgactg tgatgagcac tcaacagaat ctcgatgctg tcgttaccct ctaactgtgg
     1801 attttgaagc ttttggatgg gactggatta ttgcacccaa aagatataag gccagttact
     1861 gctctggaga gtgtgaattt gtatttttac aaaaataccc tcacactcat cttgtgcacc
     1921 aagcaaaccc cagaggttca gcaggcccct gctgtactcc cacaaagatg tctccaatca
     1981 atatgctata ttttaatggc aaagaacaaa taatatatgg gaaaattcca gccatggtag
     2041 tagatcgctg tgggtgctca ggatcccggg ctgactacaa agaccatgac ggtgattata
     2101 aagatcatga catcgactac aaggatgacg atgacaagta gtgatcccgg gtggcatccc
     2161 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     2221 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     2281 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     2341 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     2401 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     2461 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     2521 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     2581 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa
     2641 ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg
     2701 ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg
     2761 cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga
     2821 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2881 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2941 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3001 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3061 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggaattgat
     3121 cagcttggga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3181 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3241 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3301 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     3361 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3421 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3481 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3541 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3601 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3661 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     3721 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3781 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3841 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3901 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     3961 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4021 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4081 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc cgggctcgat
     4141 cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg
     4201 cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc
     4261 gaactgcagg agtggggagg cacgatggcc gctttggtcg acccggacgg gacgctcctg
     4321 cgcctgatac agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc cgttttccgc
     4381 ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag ctgctcacca
     4441 cccactcgcc aagctggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4501 gccgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     4561 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     4621 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4681 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4741 gatcaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg
     4801 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4861 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4921 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4981 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5041 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5101 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5161 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5221 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5281 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5341 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5401 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5461 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5521 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5581 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5641 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5701 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6901 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     6961 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     7021 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     7081 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     7141 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     7201 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     7261 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     7321 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     7381 cttacaattt
//
