LOCUS       .                       5621 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /label="EF-1α promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
     promoter        1196..1214
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1293..1317
                     /label="attB1"
                     /note="recombination site for the Gateway® BP reaction"
     CDS             1341..1367
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     protein_bind    complement(1387..1411)
                     /label="attB2"
                     /note="recombination site for the Gateway® BP reaction"
     CDS             1464..1505
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     polyA_signal    1549..1773
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    1843..1876
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2334..2350)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2824..2928
                     /label="AmpR promoter"
     CDS             2929..3786
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      3960..4548
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    4836..4857
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4872..4902
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4910..4926
                     /label="lac operator"
                     /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)."
     primer_bind     4934..4950
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        5623..6800
                     /label="EF-1α promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
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 tacccatacg acgtcccaga ctacgcttga aagggtgggc
     1381 gcgccgaccc agctttcttg tacaaagtgg ttgatatcca gcacagtggc ggccgctcga
     1441 gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     1501 ctacgcgtac cggttagtaa tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     1561 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1621 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1681 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1741 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1801 gctctagggg gtatccccac gcgccctgta gcggcgcatt aaataacttc gtataatgta
     1861 tgctatacga agttatatcg tatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     1921 ggactttcca caccctaact gacacacatt ccacagaatt ccttgaaata catccacagg
     1981 tacacctcca attgactcaa atgatgtcaa ttagtctatc agaagcttct aaagccatga
     2041 catcattttc tggaattttc caagctgttt aaaggcacag tcaacttagt gtatgtaaac
     2101 ttctgaccca ctggaattgt gatacagtga attataagtg aaataatctg tctgtaaaca
     2161 attgttggaa aaatgacttg tgtcatgcac aaagtagatg tcctaactga cttgccaaaa
     2221 ctattgtttg ttaacaagaa atttgtggag tagttgaaaa acgagtttta atgactccaa
     2281 cttaagtgta tgtaaacttc cgacttcaac tgtagggtac cgagctcgaa ttcactggcc
     2341 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2401 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2461 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     2521 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     2581 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2641 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2701 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     2761 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2821 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2881 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2941 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3001 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3061 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3121 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3181 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3241 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3301 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3361 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3421 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3481 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3541 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3601 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3661 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3721 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3781 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3841 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3901 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3961 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4021 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4081 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4141 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4201 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4261 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4321 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4381 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4441 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4501 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4561 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4621 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4681 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4741 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4801 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     4861 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     4921 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
     4981 gtcgactcta gacctataca gttgaagtcg gaagtttaca tacacttaag ttggagtcat
     5041 taaaactcgt ttttcaacta ctccacaaat ttcttgttaa caaacaatag ttttggcaag
     5101 tcagttagga catctacttt gtgcatgaca caagtcattt ttccaacaat tgtttacaga
     5161 cagattattt cacttataat tcactgtatc acaattccag tgggtcagaa gtttacatac
     5221 actaagttga ctgtgccttt aaacagcttg gaaaattcca gaaaatgatg tcatggcttt
     5281 agaagcttct gatagactaa ttgacatcat ttgagtcaat tggaggtgta cctgtggatg
     5341 tatttcaagg ggatccagac atgataagat acattgatga gtttggacaa accacaacta
     5401 gaatgcagcg gccgctagct cgagatctta attaaactag tatcgatata acttcgtata
     5461 atgtatgcta tacgaagtta tatgtacggg ccagatatac gcgttgacat tgattattga
     5521 ctaggctttt gcaaaaagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt
     5581 gcagctaatg gaccttctag gtcttgaaag gagtgggaat t
//
