LOCUS       sequence-39673-e        5497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5497)
  TITLE     Glazebrook/ Katagiri Lab Gateway destination vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5497)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBPX_C1
FEATURES             Location/Qualifiers
     source          1..5497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(36..464)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(146..167)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     358..377
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     591..613
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     605..622
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     606..622
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        join(715..1013,1014..1036,1037..1224)
                     /label=35SPPDK promoter
                     /note="hybrid promoter consisting of the cauliflower mosaic
                     virus 35S enhancer fused to the maize C4PPDK basal promoter
                     (Yoo et al., 2007)"
     protein_bind    1255..1379
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1404..1409,1410..1427,1428..1434)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1488..2147
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1554..1573)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2489..2794
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2673..2692
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2835..2959)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3017..3269
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(3315..3333)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3354..3370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3354..3370)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3367..3389)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3378..3394
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3402..3408,3409..3426,3427..3432))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3447..3468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3585..3602)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3756..4344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3836..3855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4515..5306,5307..5375))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5138..5157
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5376..5480)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
       61 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
      121 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
      181 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
      241 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
      301 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
      361 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
      421 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
      481 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      541 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      601 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattgggtac
      661 cgctagctac tccaagaata tcaaagatac agtctcagaa gaccaaaggg ctattgagac
      721 ttttcaacaa agggtaatat cgggaaacct cctcggattc cattgcccag ctatctgtca
      781 cttcatcaaa aggacagtag aaaaggaagg tggcacctac aaatgccatc attgcgataa
      841 aggaaaggct atcgttcaag atgcctctgc cgacagtggt cccaaagatg gacccccacc
      901 cacaaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg
      961 atgtgatatc tccactgacg taagggatga cgcacaatcc cactatcctt cgccccaagc
     1021 ttgggcccaa gcttgggtcg cgccccacgg atggtataag aataaaggca ttccgcgtgc
     1081 aggattcacc cgttcgcctc tcaccttttc gctgtactct ctcgccacac acaccccctc
     1141 tccagctccg ttggagctcc ggacagcagc aggcgcgggg cggtcacgta gtaagcagct
     1201 ctcggctccc tctccccttg ctccgtggaa tttcgaggcg cgccaagcta tcaaacaagt
     1261 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
     1321 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
     1381 cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
     1441 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     1501 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1561 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1621 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1681 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     1741 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1801 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1861 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1921 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1981 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     2041 tgccgctggc gattcaggtt catcatgccg tctgtgatgg cttccatgtc ggcagaatgc
     2101 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaatct agaggatccg
     2161 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     2221 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     2281 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     2341 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     2401 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     2461 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     2521 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     2581 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtccccc gtgaacttta
     2641 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     2701 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     2761 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     2821 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     2881 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     2941 ctttcttgta caaagtggtt cgataattcc ttaattaact agttctagag cggccgccac
     3001 cgcggtggag ctcgcagatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     3061 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     3121 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     3181 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     3241 gcgcgcggtg tcatctatgt tactagatcc gatgataagc tgtcaaacat gctagcgagc
     3301 tccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag
     3361 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3421 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3481 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3541 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3601 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3661 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3721 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3781 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3841 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3901 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3961 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4021 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4081 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4141 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4201 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4261 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4321 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4381 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4441 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4501 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4561 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4621 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4681 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4741 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4801 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4861 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4921 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4981 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5041 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5101 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5161 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5221 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5281 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5341 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5401 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5461 aaacaaatag gggttccgcg cacatttccc cgaaaag
//
