LOCUS       V028064                 6995 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028064
VERSION     V028064
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"
     protein_bind    1293..1317
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1342..2829
                     /label="Plagl2(NM_018807)"
                     /note="Plagl2(NM_018807)"
                     /gene="Plagl2"
     CDS             2830..2895
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2923..3147
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    3217..3250
                     /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(3708..3724)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        4198..4302
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4303..5163
                     /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      5334..5922
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    6210..6231
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        6246..6276
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6284..6300
                     /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     6308..6324
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6822..6855
                     /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 gtacaaaaaa gcaggctccg
     1321 cggccgcccc cttcaccatg gatgaccaca tttttcacca gcgtccctcc ctggattcaa
     1381 gatgcaaagc aggaggagga agtgggctgg aaactagttc ccaggcctcg gggccgggag
     1441 gcagagagtc aagtgaagtg ccaatgtgaa atctcgggga caccattttc aaatggggag
     1501 aagctaaggc ctcacagcct tccccatccg gagcagagac catatagctg tcctcagctg
     1561 cactgcggca aggcttttgc ctccaagtac aagctgtata ggcacatggc cacccactcg
     1621 gcccagaaac cccaccagtg catgtactgt gacaagatgt ttcaccgaaa ggaccatctg
     1681 cggaaccact tgcagaccca tgaccccaac aaagaggccc tccactgctc tgagtgtggt
     1741 aagaattata atacaaagct gggataccgt cgtcacctgg ccatgcatgc tgccagcagt
     1801 ggtgatctca gctgcaaggt gtgcctgcag acctttgaga gtacccaggc cctattggag
     1861 cacctgaagg ctcactcacg ccgggtagca ggaggtgcta aggagaagaa gcacccctgt
     1921 gaccattgtg acaggcggtt ctatactcgg aaggatgtcc gacggcacct agtggtgcac
     1981 acaggtcgta aggacttcct gtgtcagtac tgtgcccaga gatttggccg caaggatcac
     2041 ctgacacgcc atgtcaagaa gagccactca caggaactgc tcaagatcaa gacagagcca
     2101 gtggatatgt taggcttgct cagctgtagc tctacagtca gtgtaaagga agagctgagc
     2161 cctgtgcttt gcatggcctc tcgggacgtg atgggggcta aggcctttcc tggcatgttg
     2221 cccatgggta tgtatggtgc ccacatccct accatgccca gtgcgggcat gccacactcc
     2281 ctggtgcaca acacactgcc catgggtatg agttaccctc tggaatcttc tcctatctcc
     2341 tcctcagctc agctccctcc aaaataccag cttggatcta cctcatactt gcccgacaaa
     2401 ctgcccaaag tggaggtgga tagttttctg gcagagcttc ctggaagcct gtctctctcc
     2461 tcggctgaac ctcagcccgc ctcacctcag ccggcggcag ctgcagccct cctggatgaa
     2521 gcactgctcg ccaagagtcc cgccaacctc tccgaggccc tctgcgctgc taatgtggac
     2581 ttttcccact tactgggctt tcttccgctc aacctacccc catgtaaccc accaggggcc
     2641 acaggaggct tggttatggg ctactcccaa gctgaggcac agcctttgct caccactttg
     2701 caggctcagc ctcaagattc cgctggagct gggggacccc tgaactttgg gcctctgcac
     2761 tccttacctc ctgtcttcac ctctggcctg agtaccacca ccctgcctcg tttccaccaa
     2821 gcattccagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     2881 gatgacgatg acaagtagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     2941 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3001 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3061 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3121 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3181 gggggtatcc ccacgcgccc tgtagcggcg cattaaataa cttcgtataa tgtatgctat
     3241 acgaagttat atcgtatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     3301 tccacaccct aactgacaca cattccacag aattccttga aatacatcca caggtacacc
     3361 tccaattgac tcaaatgatg tcaattagtc tatcagaagc ttctaaagcc atgacatcat
     3421 tttctggaat tttccaagct gtttaaaggc acagtcaact tagtgtatgt aaacttctga
     3481 cccactggaa ttgtgataca gtgaattata agtgaaataa tctgtctgta aacaattgtt
     3541 ggaaaaatga cttgtgtcat gcacaaagta gatgtcctaa ctgacttgcc aaaactattg
     3601 tttgttaaca agaaatttgt ggagtagttg aaaaacgagt tttaatgact ccaacttaag
     3661 tgtatgtaaa cttccgactt caactgtagg gtaccgagct cgaattcact ggccgtcgtt
     3721 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3781 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3841 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3901 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3961 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4021 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4081 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4141 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4201 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4261 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4321 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4381 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4441 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4501 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4561 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4621 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4681 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4741 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4801 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4861 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4921 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4981 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5041 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5101 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5161 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5221 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5281 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5341 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5401 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5461 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5521 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5581 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5641 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5701 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5761 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5821 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5881 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5941 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6001 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6061 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6121 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6181 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     6241 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     6301 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac
     6361 tctagaccta tacagttgaa gtcggaagtt tacatacact taagttggag tcattaaaac
     6421 tcgtttttca actactccac aaatttcttg ttaacaaaca atagttttgg caagtcagtt
     6481 aggacatcta ctttgtgcat gacacaagtc atttttccaa caattgttta cagacagatt
     6541 atttcactta taattcactg tatcacaatt ccagtgggtc agaagtttac atacactaag
     6601 ttgactgtgc ctttaaacag cttggaaaat tccagaaaat gatgtcatgg ctttagaagc
     6661 ttctgataga ctaattgaca tcatttgagt caattggagg tgtacctgtg gatgtatttc
     6721 aaggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     6781 agcggccgct agctcgagat cttaattaaa ctagtatcga tataacttcg tataatgtat
     6841 gctatacgaa gttatatgta cgggccagat atacgcgttg acattgatta ttgactaggc
     6901 ttttgcaaaa agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct
     6961 aatggacctt ctaggtcttg aaaggagtgg gaatt
//
