LOCUS       V031782                 7001 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031782
VERSION     V031782
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             1348..2835
                     /label="PLAGL2(NM_002657)"
                     /note="PLAGL2(NM_002657)"
                     /gene="PLAGL2"
     CDS             2836..2901
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2929..3153
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    3223..3256
                     /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(3714..3730)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        4204..4308
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4309..5169
                     /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      5340..5928
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    6216..6237
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        6252..6282
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6290..6306
                     /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     6314..6330
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6828..6861
                     /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 ggccaccatg accacatttt tcaccagcgt ccccccctgg
     1381 attcaagatg caaagcagga ggaggaagtg ggctggaaac tagttcccag gcctcggggc
     1441 cgggaggcgg agagtcaagt gaagtgccaa tgtgaaattt cgggaacacc tttctcaaat
     1501 ggggagaagc tgaggcctca cagcctcccg caaccagagc agagaccata tagctgccct
     1561 cagctgcact gtggcaaggc ttttgcttcc aaatacaagc tgtataggca catggccacc
     1621 cactcagccc agaaacccca ccagtgtatg tactgtgata agatgtttca ccgcaaggac
     1681 catctgcgga accatctgca gacccatgat cctaacaaag aggccctcca ctgctctgag
     1741 tgcggtaaga attacaatac gaagctgggc taccggcgcc acctggccat gcatgctgcc
     1801 agcagcggtg acctcagctg caaggtgtgc ctgcagacct ttgagagtac ccaggccctg
     1861 ctagagcacc tgaaggccca ctcacgccgg gtagcaggcg gtgccaagga gaagaagcac
     1921 ccctgtgacc actgcgaccg gcggttctat actcgtaagg atgtacggcg gcacctagtg
     1981 gtgcacacag gccgtaagga cttcctgtgc cagtactgtg cccagcggtt tggccgtaag
     2041 gaccacctga cgcgtcatgt caagaagagc cactcgcagg agctgctcaa gatcaagaca
     2101 gagcccgtgg acatgttagg cctactcagc tgcagctcca cagtcagtgt gaaggaagag
     2161 ctgagccctg tgctgtgcat ggcctctcgg gacgtaatgg ggaccaaggc cttccctggc
     2221 atgttgccca tgggcatgta tggtgcccac atccctacca tgcccagcac gggcgtgcca
     2281 cactccctgg tgcacaacac gctgcccatg ggtatgagct accctctgga atcctcacct
     2341 atctcttccc cagctcagct ccctccaaaa taccagcttg gatctacctc atacttgccc
     2401 gacaaattgc ccaaagtgga ggtggatagt tttctggcgg agcttcctgg aagcctgtct
     2461 ctctcatccg ctgaacccca gcccgcctca cctcagccgg cggcagctgc ggccctccta
     2521 gatgaagcac tgcttgccaa gagccccgcc aacctctctg aggccctctg cgctgctaat
     2581 gtggacttct cccacctact gggctttctt ccactcaacc tgcccccgtg taacccacct
     2641 ggggccacag gaggcctggt catgggctac tcccaggctg aggcacagcc cctgcttacc
     2701 actttgcaag ctcagcctca agattcccca ggagctgggg gaccactgaa ctttgggcct
     2761 ctgcactcct tgcctcctgt cttcacgtct ggcctgagta gcaccaccct gcctcgtttc
     2821 catcaagcat tccaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     2881 tacaaggatg acgatgacaa gtagtttaaa cccgctgatc agcctcgact gtgccttcta
     2941 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3001 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3061 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3121 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3181 gctctagggg gtatccccac gcgccctgta gcggcgcatt aaataacttc gtataatgta
     3241 tgctatacga agttatatcg tatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     3301 ggactttcca caccctaact gacacacatt ccacagaatt ccttgaaata catccacagg
     3361 tacacctcca attgactcaa atgatgtcaa ttagtctatc agaagcttct aaagccatga
     3421 catcattttc tggaattttc caagctgttt aaaggcacag tcaacttagt gtatgtaaac
     3481 ttctgaccca ctggaattgt gatacagtga attataagtg aaataatctg tctgtaaaca
     3541 attgttggaa aaatgacttg tgtcatgcac aaagtagatg tcctaactga cttgccaaaa
     3601 ctattgtttg ttaacaagaa atttgtggag tagttgaaaa acgagtttta atgactccaa
     3661 cttaagtgta tgtaaacttc cgacttcaac tgtagggtac cgagctcgaa ttcactggcc
     3721 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3781 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3841 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     3901 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     3961 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4021 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4081 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4141 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4201 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4261 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4321 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4381 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4441 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4501 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4561 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4621 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4681 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4741 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4801 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4861 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4921 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4981 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5041 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5101 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5161 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5221 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5281 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5341 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5401 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5461 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5521 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5581 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5641 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5701 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5761 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5821 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5881 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5941 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6001 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6061 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6121 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6181 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6241 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     6301 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
     6361 gtcgactcta gacctataca gttgaagtcg gaagtttaca tacacttaag ttggagtcat
     6421 taaaactcgt ttttcaacta ctccacaaat ttcttgttaa caaacaatag ttttggcaag
     6481 tcagttagga catctacttt gtgcatgaca caagtcattt ttccaacaat tgtttacaga
     6541 cagattattt cacttataat tcactgtatc acaattccag tgggtcagaa gtttacatac
     6601 actaagttga ctgtgccttt aaacagcttg gaaaattcca gaaaatgatg tcatggcttt
     6661 agaagcttct gatagactaa ttgacatcat ttgagtcaat tggaggtgta cctgtggatg
     6721 tatttcaagg ggatccagac atgataagat acattgatga gtttggacaa accacaacta
     6781 gaatgcagcg gccgctagct cgagatctta attaaactag tatcgatata acttcgtata
     6841 atgtatgcta tacgaagtta tatgtacggg ccagatatac gcgttgacat tgattattga
     6901 ctaggctttt gcaaaaagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt
     6961 gcagctaatg gaccttctag gtcttgaaag gagtgggaat t
//
