LOCUS       sequence-229073-       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  TITLE     Petr Svoboda lab unpublished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        9..1191
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          240..1182
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1139..1159
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1223..1242
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1223..1241
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6365..6394
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6410..6427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6450..6467)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6456..6680
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7021..7040
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7160..7558
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    7716..7837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7753..7772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7807..7826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(7962..8550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8042..8061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8721..9512,9513..9581))
                     /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     9344..9363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9589..9693)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9720..10148
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9807..9826)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10017..10038
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acgcgtgagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
       61 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaagtggcgc ggggtaaact
      121 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
      181 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
      241 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
      301 cttgaattac ttccacgccc ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
      361 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
      421 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
      481 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
      541 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
      601 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
      661 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
      721 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
      781 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
      841 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
      901 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
      961 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg
     1021 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     1081 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     1141 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg acgcgtccca
     1201 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
     1261 taagcttctc tgcatcagct ctgccttcag gttctgactt ccttcgacaa tgagccgtga
     1321 tacagaagta tacacttctc agccttgtga gattgtggta gactgtggac cgtttactga
     1381 cagaagtggg ctgtatgaga gattgctgat ggagttagaa gcagcacttg attttatcaa
     1441 tgattgtaat gtagctgtac attcaaagga aagagattct actttgattt ccaaacaaat
     1501 cctctcagac tgccgtgcag tgttggttgt tctgggaccc tggtgtgcag ataaagtagc
     1561 tgggatgatg gtaagagaac ttcagaagta catcaaacat gagcaagagg agctacacag
     1621 gaagttccta ttgtttacag acaccttgtt aaggaaaata cacgccctgt gtgaagagta
     1681 cttctcacct gcctcacttg acctgaagta tgtaactccc aaagtcatga aactgctcga
     1741 gatcctgcgc aaatacaagc cctatgagcg acagcagttt gaaagtgttg agtggtacaa
     1801 taacaggaat caggataatt acgtgtcctg gagtgattcc gaggatgacg atgatgacga
     1861 agaaattgaa gaaaaagaaa agccagagac aaattttcct tctccgttta ccaatatttt
     1921 atgtgggatt atttttgtgg aaagaagata cacagcagtt gtcctaaaca gattgataaa
     1981 ggaagctggc aaacaagatc cagagctggc ttatatcagc agcaacttta taactggaca
     2041 cggcattgga aagaaccagc ctcgcagcaa acagatggag gcggagttca gaaagcagga
     2101 ggaggtactt aggaaatttc gagctcatga gaccaacctg ctcattgcaa caagtgttgt
     2161 agaagagggt gtggatattc ccaagtgcaa tttggtggtt cgttttgatt tgcctacaga
     2221 gtatcgatca tatgtccagt ctaaaggacg ggcacgagcg ccgatctcta attacgtaat
     2281 gttagcagat acagacaaga taaagagctt tgaagaagac ctcaagacct acaaagctat
     2341 tgaaaagatc ttaaggaaca agtgttcgaa gtcagcggat ggtgctgagg cggacgtcca
     2401 tgctggtgta gatgatgagg atgctttccc gccatatgtg ctgaggcctg atgatggggg
     2461 tcctcgagtc acaatcaaca ctgccattgg acacatcaac agatattgtg ctcgattacc
     2521 aagtgatccg tttacgcatc tagctcccaa atgtagaacc cgagaattgc ctgatggtac
     2581 attttattca actctttatc ttccgattaa ctcacctctt cgagcctcca ttgttggtcc
     2641 accgatggac agtgtacggt tggctgaaag agtggtagct ctcatttgct gtgaaaaact
     2701 gcacaaaatt ggtgaactgg atgaacactt gatgccagtt gggaaagaga ctgttaaata
     2761 tgaagaagag ctggacttac atgacgagga ggagaccagt gttcccggga gaccagggtc
     2821 cacaaaacga cggcagtgct accccaaagc aattccagag tgcttgaggg agagttaccc
     2881 caaacccgac cagccttgtt acctgtatgt gataggaatg gtcttaacta cccccctccc
     2941 tgatgaactc aacttcaggc ggcggaagct ctacccacct gaggacacca caagatgctt
     3001 tgggatcctg acagccaaac ccatacctca gattcctcac tttcctgtgt atacacgctc
     3061 tggagaggtc accatatcca tcgagctgaa gaagtctggt ttcacattgt ctcaacaaat
     3121 gctcgagctg attacaagat tgcaccagta catattctca catattctcc ggcttgagaa
     3181 gcctgccctg gagtttaaac ctacaggcgc ggagtcagcc tactgtgttc tacctcttaa
     3241 tgttgttaat gactccggca ctttggacat tgactttaaa ttcatggagg atattgagaa
     3301 gtcggaagct cggataggca ttcctagcac caagtattca aaggaaacac cgtttgtctt
     3361 taagttagaa gattaccaag atgcagtgat cattccaaga taccgcaatt ttgatcagcc
     3421 tcatcgattt tatgtagctg atgtatatac ggatcttacc ccactcagta aatttccttc
     3481 ccctgaatat gaaacttttg cagaatatta taaaacaaag tataaccttg acctgaccaa
     3541 tctcaaccag ccactgctgg atgttgacca cacatcttcc agactgaacc ttttgacacc
     3601 tcggcatttg aatcagaagg ggaaagcact tcctctaagc agtgctgaga agaggaaggc
     3661 caaatgggaa agtctgcaga acaaacagat cctggttccg gaactctgtg ctatacatcc
     3721 aatcccagca tcactgtgga gaaaagcggt ttgtctcccc agcatactgt atcgccttca
     3781 ctgccttctg actgcagagg aactaagagc tcagacagcc agcgatgctg gtgtgggtgt
     3841 cagatcactt cccgtggatt ttagataccc taacttagac ttcgggtgga aaaaatctat
     3901 tgatagcaag tctttcatct caagctgtaa ctcctcgttg gctgagagtg ataattactg
     3961 taagcacagc acaactgtag tccctgaaca tgctgcacat caaggtgcta ctagaccctc
     4021 cttagaaaac catgaccaaa tgtctgtgaa ctgcaaaagg ttgcccgctg agtcacctgc
     4081 taagctccaa agtgaggttt caacggatct tacagcaatt aatggtcttt cttacaataa
     4141 aaaccttgcc aatggcagtt atgatttagt taacagagac ttttgccaag gaaatcagct
     4201 gaattacttc aagcaggaaa tacctgtaca accaactacc tcatatccca ttcagaattt
     4261 gtacaattat gagaaccagc ccaagcccag caatgaatgt cctctactga gtaatacgta
     4321 ccttgatggg aacgctaaca catctacctc agatggaagc cccgcggtgt ccacgatgcc
     4381 tgccatgatg aatgctgtta aagcactcaa ggacaggatg gactctgagc agagcccttc
     4441 tgttgggtac tcctcaagga ctcttggccc caatccggga ctgattctcc aggctttgac
     4501 actgtcaaat gccagtgatg ggtttaacct ggagcggctt gagatgcttg gcgactcctt
     4561 tttaaagcat gctatcacca catatctgtt ttgcacgtac cctgatgctc atgaaggccg
     4621 cctttcatat atgagaagca aaaaggtcag caactgtaac ctgtatcgcc ttggcaagaa
     4681 gaagggactg cccagccgca tggtggtgtc gatattcgat cctcctgtga attggcttcc
     4741 tcctggttat gtggtaaacc aagacaaaag taactcagag aagtgggaaa aggatgaaat
     4801 gacaaaagac tgcttgctgg ctaatggcaa actgggtgag gcctgtgagg aggaggagga
     4861 cctgacgtgg agggccccca aggaggaggc cgaggatgag gacgacttcc tggagtatga
     4921 ccaggagcat atccagttta tagacagcat gctcatgggc tcgggcgcct tcgtgaggaa
     4981 aatctctctg tcgcccttct cagcctccga ctctgcatac gaatggaaga tgcccaagaa
     5041 ggcctccttg ggcagtatgc cgtttgcctc gggccttgag gattttgatt acagctcttg
     5101 ggatgcgatg tgctatctgg atcctagcaa agccgttgaa gaggatgact ttgtggtagg
     5161 tttctggaat ccatcagaag agaactgtgg cgtcgacaca gggaagcagt ccatttctta
     5221 cgacttgcac acggagcagt gcattgctga caagagcata gcggactgtg tggaggccct
     5281 gctgggctgc tacttaacca gctgcggcga gagggctgct cagctcttcc tctgttcgct
     5341 ggggctgaag gtgctcccag taatcaaaag gaccagccgg gagaaggcct tggaccctgc
     5401 tcaggagaac ggaagcagcc aacaaaagag cctttcaggg agctgtgcct ctccagttgg
     5461 cccacgctcc tctgcaggga aagacttgga gtacggctgc ctgaagatcc caccgaggtg
     5521 catgtttgac catccagacg cagagaaaac cctcaaccac ctgatatctg ggtttgaaac
     5581 ttttgagaag aaaattaact atagattcaa gaataaggct tatcttctgc aggcttttac
     5641 acacgcctcc taccactaca acactatcac tgattgttac cagcgcttag aattcctggg
     5701 agatgcgatt ttggactacc tcataaccaa gcacctttat gaagacccac ggcagcattc
     5761 tccaggggtc ttgactgact tgcgctctgc ccttgtcaat aacaccatct ttgcatcatt
     5821 ggctgtgaag tatgactacc acaagtactt taaagctgtc tctcctgagc tctttcatgt
     5881 tattgatgac tttgtgaagt ttcagcttga gaagaacgaa atgcaaggaa tggactctga
     5941 gcttaggaga tccgaggagg atgaggagaa agaagaggat atcgaagtcc ccaaggccat
     6001 gggggacatt tttgagtctc ttgctggtgc catttatatg gacagtggga tgtctctgga
     6061 ggtggtctgg caggtgtact atccgatgat gcagcctcta atagaaaagt tttctgcaaa
     6121 tgtgcctcgt tctccagtga gagaattgct cgagatggaa ccagaaactg ccaagtttag
     6181 cccagcggag agaacttatg atggaaaggt cagagtcacc gtggaagtgg tgggaaaggg
     6241 gaaatttaaa ggtgttggcc ggagctacag gatcgccaag tctgctgcag cacgaagggc
     6301 cctccgcagc ctcaaagcta accagcctca ggttcctaac agctcgctag agggcccgcg
     6361 gttcgaacaa aaactcatct cagaagagga tctgaatatg cataccggtc atcatcacca
     6421 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     6481 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     6541 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     6601 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     6661 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc cggtaggcgc caaccggctc
     6721 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
     6781 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc
     6841 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
     6901 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
     6961 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
     7021 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
     7081 gcctttcgac ctgcagccca agctagcatc ggcatagtat atcggcatag tataatacga
     7141 caaggtgagg aactaaacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     7201 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     7261 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     7321 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     7381 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     7441 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     7501 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagc
     7561 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     7621 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     7681 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     7741 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7801 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtatg catggcggta
     7861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8401 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8581 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8701 gagtaacctg aggctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8761 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8821 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8881 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8941 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9001 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9061 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9121 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9181 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9241 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9301 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     9361 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     9421 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     9481 gcatctttta ctttcaccag cgtttcaggg tgagcaaaaa caggaaggca aaatgccgca
     9541 aaaaagggaa taagggcgac acggaaatgt tgaatactca tcctcaggac tctccctttt
     9601 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9661 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9721 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     9781 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     9841 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     9901 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     9961 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
    10021 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
    10081 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
    10141 cgcgaatttt aacaaaatat taacgcttac aattt
//
