LOCUS       pT3-EF1a-MCL1(human)    6461 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032804
VERSION     V032804
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1196..1214
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1306..2355
                     /label="MCL1(NM_021960)"
                     /note="MCL1(NM_021960)"
                     /gene="MCL1"
     polyA_signal    2389..2613
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2683..2716
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(3174..3190)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        3664..3768
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             3769..4629
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      4800..5388
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5676..5697
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5712..5742
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5750..5766
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     primer_bind     5774..5790
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6288..6321
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaatgtt tggcctcaaa
     1321 agaaacgcgg taatcggact caacctctac tgtggggggg ccggcttggg ggccggcagc
     1381 ggcggcgcca cccgcccggg agggcgactt ttggctacgg agaaggaggc ctcggcccgg
     1441 cgagagatag ggggagggga ggccggcgcg gtgattggcg gaagcgccgg cgcaagcccc
     1501 ccgtccaccc tcacgccaga ctcccggagg gtcgcgcggc cgccgcccat tggcgccgag
     1561 gtccccgacg tcaccgcgac ccccgcgagg ctgcttttct tcgcgcccac ccgccgcgcg
     1621 gcgccgcttg aggagatgga agccccggcc gctgacgcca tcatgtcgcc cgaagaggag
     1681 ctggacgggt acgagccgga gcctctcggg aagcggccgg ctgtcctgcc gctgctggag
     1741 ttggtcgggg aatctggtaa taacaccagt acggacgggt cactaccctc gacgccgccg
     1801 ccagcagagg aggaggagga cgagttgtac cggcagtcgc tggagattat ctctcggtac
     1861 cttcgggagc aggccaccgg cgccaaggac acaaagccaa tgggcaggtc tggggccacc
     1921 agcaggaagg cgctggagac cttacgacgg gttggggatg gcgtgcagcg caaccacgag
     1981 acggccttcc aaggcatgct tcggaaactg gacatcaaaa acgaagacga tgtgaaatcg
     2041 ttgtctcgag tgatgatcca tgttttcagc gacggcgtaa caaactgggg caggattgtg
     2101 actctcattt cttttggtgc ctttgtggct aaacacttga agaccataaa ccaagaaagc
     2161 tgcatcgaac cattagcaga aagtatcaca gacgttctcg taaggacaaa acgggactgg
     2221 ctagttaaac aaagaggctg ggatgggttt gtggagttct tccatgtaga ggacctagaa
     2281 ggtggcatca ggaatgtgct gctggctttt gcaggtgttg ctggagtagg agctggtttg
     2341 gcatatctaa taagatagcc atggtttaaa cccgctgatc agcctcgact gtgccttcta
     2401 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2461 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2521 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2581 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2641 gctctagggg gtatccccac gcgccctgta gcggcgcatt aaataacttc gtataatgta
     2701 tgctatacga agttatatcg tatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     2761 ggactttcca caccctaact gacacacatt ccacagaatt ccttgaaata catccacagg
     2821 tacacctcca attgactcaa atgatgtcaa ttagtctatc agaagcttct aaagccatga
     2881 catcattttc tggaattttc caagctgttt aaaggcacag tcaacttagt gtatgtaaac
     2941 ttctgaccca ctggaattgt gatacagtga attataagtg aaataatctg tctgtaaaca
     3001 attgttggaa aaatgacttg tgtcatgcac aaagtagatg tcctaactga cttgccaaaa
     3061 ctattgtttg ttaacaagaa atttgtggag tagttgaaaa acgagtttta atgactccaa
     3121 cttaagtgta tgtaaacttc cgacttcaac tgtagggtac cgagctcgaa ttcactggcc
     3181 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3241 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3301 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     3361 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     3421 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3481 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3541 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3601 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3661 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3721 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3781 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3841 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3901 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3961 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4021 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4081 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4141 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4201 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4261 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4321 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4381 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4441 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4501 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4561 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4621 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4681 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4741 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4801 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4861 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4921 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4981 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5041 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5101 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5161 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5221 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5281 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5341 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5401 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5461 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5521 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     5581 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     5641 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     5701 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     5761 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
     5821 gtcgactcta gacctataca gttgaagtcg gaagtttaca tacacttaag ttggagtcat
     5881 taaaactcgt ttttcaacta ctccacaaat ttcttgttaa caaacaatag ttttggcaag
     5941 tcagttagga catctacttt gtgcatgaca caagtcattt ttccaacaat tgtttacaga
     6001 cagattattt cacttataat tcactgtatc acaattccag tgggtcagaa gtttacatac
     6061 actaagttga ctgtgccttt aaacagcttg gaaaattcca gaaaatgatg tcatggcttt
     6121 agaagcttct gatagactaa ttgacatcat ttgagtcaat tggaggtgta cctgtggatg
     6181 tatttcaagg ggatccagac atgataagat acattgatga gtttggacaa accacaacta
     6241 gaatgcagcg gccgctagct cgagatctta attaaactag tatcgatata acttcgtata
     6301 atgtatgcta tacgaagtta tatgtacggg ccagatatac gcgttgacat tgattattga
     6361 ctaggctttt gcaaaaagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt
     6421 gcagctaatg gaccttctag gtcttgaaag gagtgggaat t
//
