LOCUS       Exported                6386 bp ds-DNA     circular SYN 12-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Oct 16, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     GENERAL Depositing Lab: Pawel Pelczar Publication: Hermann et al 
            Nucleic Acids Res. 2014 Jan 9. Purpose: Component of the Bxb1-rcCre 
            binary site-specific recombinase system, expression of the 
            site-specific recombinase Bxb1 (mouse codon-optimized) BACKBONE 
            Backbone manufacturer: CLONTECH Backbone size w/o insert (bp): 4814 
            Vector backbone: pDIRECT Vector type: Mammalian Expression CLONING 
            INFORMATION 3′ cloning site: Esp3I (destroyed during cloning) 3′ 
            sequencing primer: GGAAAGGACAGTGGGAGTGGC 5′ cloning site: Esp3I 
            (destroyed during cloning) 5′ sequencing primer: 
            CATGCCTTCTTCTTTTTCCTAC Cloning method: Restriction Enzyme GROWTH IN 
            BACTERIA Bacterial Resistance(s): Ampicillin Copy number: High Copy 
            Growth Strain(s): DH5alpha Growth Temperature: 37(o)C GENE/INSERT 
            Gene/Insert name: NLS-HA-Bxb1 Promoter: CAG
FEATURES             Location/Qualifiers
     source          1..6386
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          join(145..6386,1..144)
                     /label=source
     protein_bind    16..37
                     /label=CAP binding site
     promoter        52..82
                     /label=lac promoter
     protein_bind    90..106
                     /label=lac operator
     primer_bind     95..117
                     /label=M13/pUC Reverse
     primer_bind     114..130
                     /label=M13 Reverse
     primer_bind     114..130
                     /label=M13 rev
     enhancer        150..529
                     /label=CMV enhancer
     promoter        531..808
                     /label=chicken beta-actin promoter
     intron          809..1825
                     /label=chimeric intron
     primer_bind     1749..1771
                     /label=pCAGGS-5
     primer_bind     1819..1861
                     /label=KJ1523
     misc_feature    1828..1863
                     /label=F common for Bxb1 mutants
     primer_bind     1838..1857
                     /label=T7
     promoter        1838..1856
                     /label=T7 promoter
     misc_feature    1841..1872
                     /label=5' Gibson
     misc_feature    1842..1865
                     /label=F primer
     CDS             1876..1896
                     /label=SV40 NLS
     misc_feature    1886..1910
                     /label=F binding
     misc_feature    1887..1907
                     /label=PD3236
     CDS             1897..1923
                     /label=HA
     CDS             1930..3429
                     /codon_start=1
                     /label=bxb1-start
                     /translation="RALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLDV
                     SGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLVV
                     SATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGYL
                     PTRVDGEWRLVPDPVQRERILEVYHRVVDGHEPLHLVAHDLNRRGVLSPKDYFAQLQGR
                     EPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAEL
                     VKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTVA
                     MAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAGS
                     PQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSMN
                     VRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     primer_bind     2140..2173
                     /label=Primer 2
     primer_bind     complement(2226..2253)
                     /label=KJ1522
     primer_bind     complement(2594..2631)
                     /label=Primer 1
     primer_bind     2614..2643
                     /label=Primer 4
     primer_bind     complement(3034..3069)
                     /label=Primer 6
     primer_bind     3052..3075
                     /label=Primer 5
     misc_feature    complement(3403..3426)
                     /label=PD3237
     primer_bind     complement(3420..3446)
                     /label=KJ1534
     misc_feature    3430..3465
                     /label=3' Gibson
     primer_bind     complement(3449..3466)
                     /label=BGH-rev
     polyA_signal    3455..3679
                     /label=bGH poly(A) signal
     misc_feature    3724..3749
                     /label=F 191
     primer_bind     complement(3751..3768)
                     /label=M13 Forward
     primer_bind     complement(3751..3767)
                     /label=M13 fwd
     primer_bind     complement(3760..3782)
                     /label=M13/pUC Forward
     rep_origin      3909..4337
                     /label=f1 ori
     primer_bind     complement(3996..4015)
                     /label=F1ori-R
     primer_bind     4206..4227
                     /label=F1ori-F
     promoter        4390..4494
                     /label=AmpR promoter
     CDS             4495..5355
                     /label=AmpR
     primer_bind     complement(4713..4732)
                     /label=Amp-R
     rep_origin      5526..6114
                     /label=ori
     primer_bind     6015..6034
                     /label=pBR322ori-F
     primer_bind     6268..6285
                     /label=L4440
ORIGIN
        1 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
       61 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      121 cagctatgac catgaggcgc gccggattcg acattgatta ttgactagtt attaatagta
      181 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      241 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      301 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      361 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      421 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      481 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
      541 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
      601 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc
      661 rggsggggsg gggsggggsg rggggsgggg sggggsgagg cggagaggtg cggcggcagc
      721 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
      781 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
      841 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
      901 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
      961 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
     1021 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     1081 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     1141 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
     1201 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1261 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
     1321 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
     1381 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga
     1441 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1501 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
     1561 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
     1621 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     1681 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     1741 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     1801 tcatgccttc ttctttttcc tacagatcct taattaataa tacgactcac tatagggggt
     1861 cgacccgcca ccatgccaaa aaagaaaaga aaagtgtatc cctatgatgt ccccgattat
     1921 gccggttcaa gagccctggt cgtgattaga ctgagccgag tgacagacgc caccacaagt
     1981 cccgagagac agctggaatc atgccagcag ctctgtgctc agcggggttg ggatgtggtc
     2041 ggcgtggcag aggatctgga cgtgagcggg gccgtcgatc cattcgacag aaagaggagg
     2101 cccaacctgg caagatggct cgctttcgag gaacagccct ttgatgtgat cgtcgcctac
     2161 agagtggacc ggctgacccg ctcaattcga catctccagc agctggtgca ttgggctgag
     2221 gaccacaaga aactggtggt cagcgcaaca gaagcccact tcgatactac cacacctttt
     2281 gccgctgtgg tcatcgcact gatgggcact gtggcccaga tggagctcga agctatcaag
     2341 gagcgaaaca ggagcgcagc ccatttcaat attagggccg gtaaatacag aggctccctg
     2401 cccccttggg gatatctccc taccagggtg gatggggagt ggagactggt gccagacccc
     2461 gtccagagag agcggattct ggaagtgtac cacagagtgg tcgatggcca cgaaccactc
     2521 catctggtgg cacacgacct gaatagacgc ggcgtgctct ctccaaagga ttattttgct
     2581 cagctgcagg gaagagagcc acagggaaga gaatggagtg ctactgcact gaagagatct
     2641 atgatcagtg aggctatgct gggttacgca acactcaatg gcaaaactgt ccgggacgat
     2701 gacggagccc ctctggtgag ggctgagcct attctcacca gagagcagct cgaagctctg
     2761 cgggcagaac tggtcaagac tagtcgcgcc aaacctgccg tgagcacccc aagcctgctc
     2821 ctgagggtgc tgttctgcgc cgtctgtgga gagccagcat acaagtttgc cggcggaggg
     2881 cgcaaacatc cccgctatcg atgcaggagc atggggttcc ctaagcactg tggaaacggg
     2941 acagtggcca tggctgagtg ggacgccttt tgcgaggaac aggtgctgga tctcctgggt
     3001 gacgctgagc ggctggaaaa agtgtgggtg gcaggatctg actccgctgt ggagctggca
     3061 gaagtcaatg ccgagctcgt ggatctgact tccctcatcg gatctcctgc atatagagct
     3121 gggtccccac agagagaagc tctggacgca cgaattgctg cactcgctgc tagacaggag
     3181 gaactggagg gcctggaggc caggccctct ggatgggagt ggcgagaaac cggacagagg
     3241 tttggggatt ggtggaggga gcaggacacc gcagccaaga acacatggct gagatccatg
     3301 aatgtccggc tcacattcga cgtgcgcggt ggcctgactc gaaccatcga ttttggcgac
     3361 ctgcaggagt atgaacagca cctgagactg gggtccgtgg tcgaaagact gcacactggg
     3421 atgtcctagg tcagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3481 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3541 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3601 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3661 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gagatccact
     3721 agttctagcc tcgaggctag agcggccgcc actggccgtc gttttacaac gtcgtgactg
     3781 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3841 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3901 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3961 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4021 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4081 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4141 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4201 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4261 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4321 aaaatttaac gcgaatttta acaaaatatt aacgcttacr mktymsrtks smcwttymgg
     4381 sgaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4441 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4501 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4561 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4621 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4681 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4741 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4801 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4861 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4921 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4981 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5041 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5101 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5161 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5221 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5281 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5341 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5641 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6181 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6241 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6301 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6361 aggtttcccg actggaaagc gggcag
//
