LOCUS       sequence-9484-ex        3580 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 3580)
  AUTHORS   Haudenshield JS, Hartman GL
  TITLE     Exogenous Controls Increase Negative Call Veracity in Multiplexed, 
            Quantitative PCR Assays for Phakopsora pachyrhizi.
  JOURNAL   Plant Dis. 2011 Mar;95(3):343-352. doi: 10.1094/PDIS-01-10-0023.
  PUBMED    30743502
REFERENCE   2  (bases 1 to 3580)
  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"; doi: "10"; journalName: 
            "Plant Dis"; date: "2011-03"; volume: "95"; issue: "3"; pages: 
            "343-352"
COMMENT     Sequence Label: pJSH-B14
FEATURES             Location/Qualifiers
     source          1..3580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..44
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        26..44
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     73..89
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     74..90
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             231..242
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(271..287)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(271..287)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(316..335)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(317..335)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(345..362)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(345..361)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(354..376)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(join(573..1364,1365..1433))
                     /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     1196..1215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1434..1538)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1608..1966
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1695..1714)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1905..1926
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    complement(1998..2031)
                     /label=lox71/66
                     /bound_moiety="Cre recombinase"
                     /note="loxP double mutant with a low affinity for Cre 
                     recombinase (Araki et al., 2010). 8-bp core sequence 
                     (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      2595..3183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3084..3103
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3337..3354
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3471..3492
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3507..3512,3513..3530,3531..3537)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3545..3561
                     /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)."
     primer_bind     3550..3572
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3569..3580,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3569..3580,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctgggta
       61 ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatccactgt ggaattcgcc
      121 cttcacgcct aggacgagaa ctcccacatc gagcttgacg caaacgacca cgccaggacc
      181 atggcaaaca tcaacaccga gcgcaacgcc ttgtgctgaa acacccgctg attgaagggc
      241 gaattccaca ttggtcgctg cagcccgggg gatccactag ttctagagcg gccgcaccgc
      301 gggagctcca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
      361 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      421 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg attaaatttt ggtcatgaga
      481 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      541 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      601 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
      661 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
      721 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      781 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      841 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      901 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      961 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1021 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1081 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1141 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1201 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1261 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1321 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1381 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1441 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1501 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1561 ccttaatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     1621 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1681 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1741 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1801 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1861 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1921 aactggaaca acactcaacc ctatctcggt ctattctttt gatttacagt taattaaagg
     1981 gaacaaaagc tggcatgtac cgttcgtata gcatacatta tacgaacggt acgctccaat
     2041 tcgcccttta attaactgtt ccaactttca ccataatgaa ataagatcac taccgggcgt
     2101 attttttgag ttgtcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     2161 tggatatacc accgagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
     2221 ttattggtgc ccttaaacgc ctggttgcta cgcctgaata agtgataata agcggatgaa
     2281 tggcagaaat tcgaaagcaa attcgacccg gtcgtcggtt cagggcaggg tcgttaaata
     2341 gccgcttatg tctattgctg gtttaccggt ttattgacta ccggaagcag tgtgaccgtg
     2401 tgcttctcaa atgcctgagg ccagtttgct caggctctcc ccgtggaggt aataattgac
     2461 gatatgatcc tttttttctg atcaaaaagg atctaggtga agatcctttt tgataatctc
     2521 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2581 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2641 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2701 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2761 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2821 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2881 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2941 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3001 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3061 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3121 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3181 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3241 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3301 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3361 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3421 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     3481 tagctcactc attaggcacc ccaggcttta cactttatgc tcccggctcg tatgttgtgt
     3541 ggaattgtga gcggataaca atttcacaca ggaaacagct
//
