LOCUS       sequence-244344-       10175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10175)
  TITLE     Petr Svoboda lab unpublished
REFERENCE   2  (bases 1 to 10175)
  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: pEF1-MH.Bl-mDcrOO
FEATURES             Location/Qualifiers
     source          1..10175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..1241
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          290..1232
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1189..1209
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1273..1292
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1273..1291
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6415..6444
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6460..6477
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6500..6517)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6506..6730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7071..7090
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7210..7608
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    7766..7887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7803..7822)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7857..7876
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(8012..8600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8092..8111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8771..9562,9563..9631))
                     /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     9394..9413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9639..9743)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(9770..10175,1..50)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9857..9876)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10067..10088
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt acgcgtgagg
       61 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
      121 aggggtcggc aattgaaccg gtgcctagag aaagtggcgc ggggtaaact gggaaagtga
      181 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
      241 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt
      301 gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac
      361 ttccacgccc ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      421 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      481 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      541 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      601 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      661 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      721 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      781 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
      841 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
      901 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
      961 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1021 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1081 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1141 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1201 cagtggttca aagttttttt cttccatttc aggtgtcgtg acgcgtccca ctgcttactg
     1261 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt taagcttctc
     1321 tgcatcagct ctgccttcag gttctgactt ccttcgacaa tgagccgtga tacagaagta
     1381 tacacttctc agccttgtga gattgtggta gactgtggac cgtttactga cagaagtggg
     1441 ctgtatgaga gattgctgat ggagttagaa gcagcacttg attttatcaa tgattgtaat
     1501 gtagctgtac attcaaagga aagagattct actttgattt ccaaacaaat cctctcagac
     1561 tgccgtgcag tgttggttgt tctgggaccc tggtgtgcag ataaagtagc tgggatgatg
     1621 gtaagagaac ttcagaagta catcaaacat gagcaagagg agctacacag gaagttccta
     1681 ttgtttacag acaccttgtt aaggaaaata cacgccctgt gtgaagagta cttctcacct
     1741 gcctcacttg acctgaagta tgtaactccc aaagtcatga aactgctcga gatcctgcgc
     1801 aaatacaagc cctatgagcg acagcagttt gaaagtgttg agtggtacaa taacaggaat
     1861 caggataatt acgtgtcctg gagtgattcc gaggatgacg atgatgacga agaaattgaa
     1921 gaaaaagaaa agccagagac aaattttcct tctccgttta ccaatatttt atgtgggatt
     1981 atttttgtgg aaagaagata cacagcagtt gtcctaaaca gattgataaa ggaagctggc
     2041 aaacaagatc cagagctggc ttatatcagc agcaacttta taactggaca cggcattgga
     2101 aagaaccagc ctcgcagcaa acagatggag gcggagttca gaaagcagga ggaggtactt
     2161 aggaaatttc gagctcatga gaccaacctg ctcattgcaa caagtgttgt agaagagggt
     2221 gtggatattc ccaagtgcaa tttggtggtt cgttttgatt tgcctacaga gtatcgatca
     2281 tatgtccagt ctaaaggacg ggcacgagcg ccgatctcta attacgtaat gttagcagat
     2341 acagacaaga taaagagctt tgaagaagac ctcaagacct acaaagctat tgaaaagatc
     2401 ttaaggaaca agtgttcgaa gtcagcggat ggtgctgagg cggacgtcca tgctggtgta
     2461 gatgatgagg atgctttccc gccatatgtg ctgaggcctg atgatggggg tcctcgagtc
     2521 acaatcaaca ctgccattgg acacatcaac agatattgtg ctcgattacc aagtgatccg
     2581 tttacgcatc tagctcccaa atgtagaacc cgagaattgc ctgatggtac attttattca
     2641 actctttatc ttccgattaa ctcacctctt cgagcctcca ttgttggtcc accgatggac
     2701 agtgtacggt tggctgaaag agtggtagct ctcatttgct gtgaaaaact gcacaaaatt
     2761 ggtgaactgg atgaacactt gatgccagtt gggaaagaga ctgttaaata tgaagaagag
     2821 ctggacttac atgacgagga ggagaccagt gttcccggga gaccagggtc cacaaaacga
     2881 cggcagtgct accccaaagc aattccagag tgcttgaggg agagttaccc caaacccgac
     2941 cagccttgtt acctgtatgt gataggaatg gtcttaacta cccccctccc tgatgaactc
     3001 aacttcaggc ggcggaagct ctacccacct gaggacacca caagatgctt tgggatcctg
     3061 acagccaaac ccatacctca gattcctcac tttcctgtgt atacacgctc tggagaggtc
     3121 accatatcca tcgagctgaa gaagtctggt ttcacattgt ctcaacaaat gctcgagctg
     3181 attacaagat tgcaccagta catattctca catattctcc ggcttgagaa gcctgccctg
     3241 gagtttaaac ctacaggcgc ggagtcagcc tactgtgttc tacctcttaa tgttgttaat
     3301 gactccggca ctttggacat tgactttaaa ttcatggagg atattgagaa gtcggaagct
     3361 cggataggca ttcctagcac caagtattca aaggaaacac cgtttgtctt taagttagaa
     3421 gattaccaag atgcagtgat cattccaaga taccgcaatt ttgatcagcc tcatcgattt
     3481 tatgtagctg atgtatatac ggatcttacc ccactcagta aatttccttc ccctgaatat
     3541 gaaacttttg cagaatatta taaaacaaag tataaccttg acctgaccaa tctcaaccag
     3601 ccactgctgg atgttgacca cacatcttcc agactgaacc ttttgacacc tcggcatttg
     3661 aatcagaagg ggaaagcact tcctctaagc agtgctgaga agaggaaggc caaatgggaa
     3721 agtctgcaga acaaacagat cctggttccg gaactctgtg ctatacatcc aatcccagca
     3781 tcactgtgga gaaaagcggt ttgtctcccc agcatactgt atcgccttca ctgccttctg
     3841 actgcagagg aactaagagc tcagacagcc agcgatgctg gtgtgggtgt cagatcactt
     3901 cccgtggatt ttagataccc taacttagac ttcgggtgga aaaaatctat tgatagcaag
     3961 tctttcatct caagctgtaa ctcctcgttg gctgagagtg ataattactg taagcacagc
     4021 acaactgtag tccctgaaca tgctgcacat caaggtgcta ctagaccctc cttagaaaac
     4081 catgaccaaa tgtctgtgaa ctgcaaaagg ttgcccgctg agtcacctgc taagctccaa
     4141 agtgaggttt caacggatct tacagcaatt aatggtcttt cttacaataa aaaccttgcc
     4201 aatggcagtt atgatttagt taacagagac ttttgccaag gaaatcagct gaattacttc
     4261 aagcaggaaa tacctgtaca accaactacc tcatatccca ttcagaattt gtacaattat
     4321 gagaaccagc ccaagcccag caatgaatgt cctctactga gtaatacgta ccttgatggg
     4381 aacgctaaca catctacctc agatggaagc cccgcggtgt ccacgatgcc tgccatgatg
     4441 aatgctgtta aagcactcaa ggacaggatg gactctgagc agagcccttc tgttgggtac
     4501 tcctcaagga ctcttggccc caatccggga ctgattctcc aggctttgac actgtcaaat
     4561 gccagtgatg ggtttaacct ggagcggctt gagatgcttg gcgactcctt tttaaagcat
     4621 gctatcacca catatctgtt ttgcacgtac cctgatgctc atgaaggccg cctttcatat
     4681 atgagaagca aaaaggtcag caactgtaac ctgtatcgcc ttggcaagaa gaagggactg
     4741 cccagccgca tggtggtgtc gatattcgat cctcctgtga attggcttcc tcctggttat
     4801 gtggtaaacc aagacaaaag taactcagag aagtgggaaa aggatgaaat gacaaaagac
     4861 tgcttgctgg ctaatggcaa actgggtgag gcctgtgagg aggaggagga cctgacgtgg
     4921 agggccccca aggaggaggc cgaggatgag gacgacttcc tggagtatga ccaggagcat
     4981 atccagttta tagacagcat gctcatgggc tcgggcgcct tcgtgaggaa aatctctctg
     5041 tcgcccttct cagcctccga ctctgcatac gaatggaaga tgcccaagaa ggcctccttg
     5101 ggcagtatgc cgtttgcctc gggccttgag gattttgatt acagctcttg ggatgcgatg
     5161 tgctatctgg atcctagcaa agccgttgaa gaggatgact ttgtggtagg tttctggaat
     5221 ccatcagaag agaactgtgg cgtcgacaca gggaagcagt ccatttctta cgacttgcac
     5281 acggagcagt gcattgctga caagagcata gcggactgtg tggaggccct gctgggctgc
     5341 tacttaacca gctgcggcga gagggctgct cagctcttcc tctgttcgct ggggctgaag
     5401 gtgctcccag taatcaaaag gaccagccgg gagaaggcct tggaccctgc tcaggagaac
     5461 ggaagcagcc aacaaaagag cctttcaggg agctgtgcct ctccagttgg cccacgctcc
     5521 tctgcaggga aagacttgga gtacggctgc ctgaagatcc caccgaggtg catgtttgac
     5581 catccagacg cagagaaaac cctcaaccac ctgatatctg ggtttgaaac ttttgagaag
     5641 aaaattaact atagattcaa gaataaggct tatcttctgc aggcttttac acacgcctcc
     5701 taccactaca acactatcac tgattgttac cagcgcttag aattcctggg agatgcgatt
     5761 ttggactacc tcataaccaa gcacctttat gaagacccac ggcagcattc tccaggggtc
     5821 ttgactgact tgcgctctgc ccttgtcaat aacaccatct ttgcatcatt ggctgtgaag
     5881 tatgactacc acaagtactt taaagctgtc tctcctgagc tctttcatgt tattgatgac
     5941 tttgtgaagt ttcagcttga gaagaacgaa atgcaaggaa tggactctga gcttaggaga
     6001 tccgaggagg atgaggagaa agaagaggat atcgaagtcc ccaaggccat gggggacatt
     6061 tttgagtctc ttgctggtgc catttatatg gacagtggga tgtctctgga ggtggtctgg
     6121 caggtgtact atccgatgat gcagcctcta atagaaaagt tttctgcaaa tgtgcctcgt
     6181 tctccagtga gagaattgct cgagatggaa ccagaaactg ccaagtttag cccagcggag
     6241 agaacttatg atggaaaggt cagagtcacc gtggaagtgg tgggaaaggg gaaatttaaa
     6301 ggtgttggcc ggagctacag gatcgccaag tctgctgcag cacgaagggc cctccgcagc
     6361 ctcaaagcta accagcctca ggttcctaac agctcgctag agggcccgcg gttcgaacaa
     6421 aaactcatct cagaagagga tctgaatatg cataccggtc atcatcacca tcaccattga
     6481 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     6541 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     6601 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     6661 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     6721 ggctctatgg cttctgaggc ggaaagaacc cggtaggcgc caaccggctc cgttctttgg
     6781 tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca
     6841 gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat
     6901 ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct
     6961 ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag
     7021 gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac
     7081 gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac
     7141 ctgcagccca agctagcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     7201 aactaaacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     7261 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     7321 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     7381 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     7441 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     7501 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     7561 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagc acttcgtggc
     7621 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     7681 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7741 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7801 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7861 tttgtccaaa ctcatcaatg tatcttatca tgtctgtatg catggtggta atacggttat
     7921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     8041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     8101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     8161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     8221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     8281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     8341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     8401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     8461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     8521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     8581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     8701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg
     8761 aggctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     9001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     9061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     9121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     9181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     9241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     9301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     9361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     9421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     9481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     9541 ctttcaccag cgtttcaggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     9601 taagggcgac acggaaatgt tgaatactca tcctcaggac tctccctttt tcaatattat
     9661 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9721 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     9781 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     9841 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     9901 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     9961 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
    10021 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
    10081 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
    10141 ccgatttcgg cctattggtt aaaaaatgag ctgat
//
