LOCUS       sequence-200881-        7051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7051)
  AUTHORS   Sabourin LA, Rudnicki MA
  TITLE     The molecular regulation of myogenesis.
  JOURNAL   Clin Genet. 2000 Jan . 57(1):16-25.
  PUBMED    10733231
REFERENCE   2  (bases 1 to 7051)
  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: "Clin Genet.
            2000 Jan . 57(1):16-25."
COMMENT     Sequence Label: EMSV-myogenin (puro)
FEATURES             Location/Qualifiers
     source          1..7051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    92..113
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(128..133,134..151,152..158)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    166..182
                     /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)."
     primer_bind     171..193
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     190..206
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     190..206
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        227..245
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    2149..2283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2173..2192)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2227..2246
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2964..3563)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3047..3067)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3544..3563
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3603..4102)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3651..3670)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     4044..4067
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(4161..4180)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4162..4180)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4187..4204)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4187..4203)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4196..4218)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4344..4799
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4431..4450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4641..4662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4866..4885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4985..5007
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5045..5063)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5131..5235
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5236..5304,5305..6096)
                     /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(5454..5473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6267..6855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6756..6775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7009..7026
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
       61 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      121 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      181 aatttcacac aggaaacagc tatgaccatg attacgccaa gctcgaaatt aaccctcact
      241 aaagggaaca aaagcttgga aaagttcaga tcaaggtcag gaacaaagaa acagctgaat
      301 accaaacagg atatctgtgg taagcggttc ctgccccggc tcagggccaa gaacagatga
      361 gacagctgag tgatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcgg
      421 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gtgaatcatc
      481 agatgtttcc agggtgcccc aaggacctga aaatgaccct gtaccttatt tgaactaacc
      541 aatcagttcg cttctcgctt ctgttcgcgc gcttccgctc tccgagctca ataaaagagc
      601 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccc agatccccgg
      661 gcgagctcga attccggcca gtggaaggaa caagtctttt ccgacctgat ggagctgtat
      721 gaaacatccc cctatttcta ccaggagccc cacttctatg acggggaaaa ctaccttcct
      781 gtccaccttc agggcttcga gcccccgggc tatgagcgga ccgagctcag cttaagcccg
      841 gaagcccgag ggcccctaga agaaaaggga ctggggaccc ctgagcattg tccaggccag
      901 tgcctgccct gggcgtgtaa ggtgtgtaag aggaagtctg tgtctgtgga ccggcggagg
      961 gcagccacac tgagggagaa gcgcaggctc aagaaagtga atgaggcctt cgaggctctg
     1021 aagagaagca ccctgctcaa ccccaaccag cggctgccta aggtggagat cctgcgcagt
     1081 gccatccagt acattgagcg cctacaggcc ttgctcagct ccctcaacca ggaggagcgc
     1141 gatctccgct accgaggcgg gggcgggccc cagccggtgg tacccagtga atgcaactcc
     1201 cacagcgcct cctgcagtcc ggagtggggc aatgcactgg agtttggtcc caacccagga
     1261 gatcatttgc tcgcagctga ccctacaggt gcccacaacc tgcactccct tacgtccatc
     1321 gtggacagca tcacggtgga ggatatgtct gtcaccttcc cagatgaaac catgcccaac
     1381 tgagattgtc tgccaggctg ggtgtgcatg ggagcccctg agttggtgtc agacgccacc
     1441 acttctgcag caggggcctt ttaagtgggg ctgtcctggt gcccagaaaa cagccctggg
     1501 ctgccacaag ccagactacc caccgtccat tcacataaga ctaacaccca gcccagcaag
     1561 ggaatttaac tgactcctta aagcagagag tatccccttc caaggagaga aagatggagt
     1621 ccagagagcc cccttgttaa tgcccctcaa cggggcaaac tcaggagctt cctttttttt
     1681 ttcttcataa tgcgcctcta attccacccc ccaaatgaga cagtttgaga gacagtgagt
     1741 gccccgaccg ggacaagtgt gcacatctgt tcgactctct tcctgatgcc agtggctggg
     1801 ctgggcctgc cctgagatga gagagaaggg agggaacgat agggttccaa agtctctggg
     1861 gggcaagcat ttgcagtgga tattgggaac cttccagtgc tttgtgtatt gtttattgtt
     1921 gtgtgtgttg tttgtaaagc tgtcatctga ccaaggtctc ctgtgctgat gccagggaca
     1981 ggcagggaag ggggtggggg ctcttggggt gacttctttt gttaactaag cattgtgtgg
     2041 ttttgccaat tttttttgta atttttttgc taacttattt ggatttcctt ttttaaaaaa
     2101 tgaataaaga ccggttgcta tcaaaaaagg aattcgagct cgcccgggga tccagacatg
     2161 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2221 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2281 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2341 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc tctagagtcg
     2401 agcccaagct tgcatgcctg caggtcgacc agcttctgat ggaattagaa cttggcaaaa
     2461 caatactgag aatgaagtgt atgtggaaca gaggctgctg atctcgttct tcaggctatg
     2521 aaactgacac atttggaaac cacagtactt agaaccacaa agtgggaatc aagagaaaaa
     2581 caatgatccc acgagagatc tatagatcta tagatcatga gtgggaggaa tgagctggcc
     2641 cttaatttgg ttttgcttgt ttaaattatg atatccaact atgaaacatt atcataaagc
     2701 aatagtaaag agccttcagt aaagagcagg catttatcta atcccacccc acccccaccc
     2761 ccgtagctcc aatccttcca ttcaaaatgt aggtactctg ttctcaccct tcttaacaaa
     2821 gtatgacagg aaaaacttcc attttagtgg acatctttat tgtttaatag atcatcaatt
     2881 tctgcatccc ggggatctga tatcatcgat gcatggggtc gtgcgctcct ttcggtcggg
     2941 cgctgcgggt cgtggggcgg gcgtcaggca ccgggcttgc gggtcatgca ccaggtgcgc
     3001 ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg
     3061 aggttgcggg gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc
     3121 actccgggga gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgagacgccg
     3181 acggtggcca ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc
     3241 tgttgctgcg cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg
     3301 aacaccgccc ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg
     3361 tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc
     3421 tcggtgaccc gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg
     3481 gcgaacgcgg cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc
     3541 accgtgggct tgtactcggt catggtaagc ttcagctgct cgagatctag atggatgcag
     3601 gtcgaaaggc ccggagatga ggaagaggag aacagcgcgg cagacgtgcg cttttgaagc
     3661 gtgcagaatg ccgggcctcc ggaggacctt cgggcgcccg ccccgcccct gagcccgccc
     3721 ctgagcccgc ccccggaccc accccttccc agcctctgag cccagaaagc gaaggagcaa
     3781 agctgctatt ggccgctgcc ccaaaggcct acccgcttcc attgctcagc ggtgctgtcc
     3841 atctgcacga gactagtgag acgtgctact tccatttgtc acgtcctgca cgacgcgagc
     3901 tgcggggcgg gggggaactt cctgactagg ggaggagtag aaggtggcgc gaaggggcca
     3961 ccaaagaacg gagccggttg gcgcctaccg gtggatgtgg aatgtgtgcg aggccagagg
     4021 ccacttgtgt agcgccaagt gcccagcggg gctgctaaag cgcatgctcc agactgcctt
     4081 gggaaaagcg cctcccctac ccggtagaat ttcgaggtcg actctagagg atccccgggt
     4141 accgagctcg aattaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt
     4201 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4261 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4321 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4381 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4441 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4501 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4561 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4621 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4681 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4741 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     4801 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     4861 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4921 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4981 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5041 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5101 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5161 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5221 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5281 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5341 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5401 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5461 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5521 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5581 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5641 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5701 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5761 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5821 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5881 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5941 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6001 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6061 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6121 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6181 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6241 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6301 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6361 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6421 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6481 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6541 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6601 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6661 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6721 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6781 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6841 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6901 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6961 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7021 agcgaggaag cggaagagcg cccaatacgc a
//
