hhhhhhhhhhhhh Solution Gibson

June 30, 2025, 6:38 p.m.

  • 4 Parts 8 Primers
  • 60.72C Average Primer Tm 34nt Average Primer
  • 93.00% BLAST nt 7.00% Synthetic nt 3 BLAST Parts 1 Synthetic Parts 717nt Longest Part 112nt Shortest Part
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Parts Download Parts

Below is a summary table of the DNA sequences that make up this assembly solution. Further details on each part can be viewed by clicking View Part for each listed below this table.

Query Start/End: The start/end index of the part on the insert sequence. Defaults to 0.
Subject Start/End: The start/end index of the part on its original sequence. Defaults to 0.
Position: The location of the part in the assembly solution.
Name Database Length Extended Length Query Start Query End Subject Start Subject End Position
120714-addgene addgene 133 163 0 133 21069 20937 0
74625-addgene addgene 711 741 133 844 2063 2773 1
Synthetic-1.112 NONE 112 142 844 956 0 0 2
71655-addgene addgene 717 747 956 1673 6958 7674 3
pBR322_b NONE 4361 4391 1674 6034 0 0 4

Plasmid Map

Parts List

120714-addgene addgene View Part

163nt Extended 133nt Template

  • Assembly Sequence: TCTCATGTTTGACAGgtgagctgat...ctggcacgacatggtgagcaagggc
  • Template Sequence: gtgagctgataccgctcgccgcagc...gattcattaatgcagctggcacgac

74625-addgene addgene View Part

741nt Extended 711nt Template

  • Assembly Sequence: tgcagctggcacgacatggtgagca...gtacaagtaaggtgataccagcatc
  • Template Sequence: atggtgagcaagggcgaggacgaca...cggcatggacgagctgtacaagtaa

Synthetic-1.112 View Part

142nt Extended 112nt Template

  • Assembly Sequence: gagctgtacaagtaaggtgatacca...cttgttgaatatgagtaaaggagaa
  • Template Sequence: ggtgataccagcatcgtcttgatgc...ggagttgtcccaattcttgttgaat

71655-addgene addgene View Part

747nt Extended 717nt Template

  • Assembly Sequence: caattcttgttgaatatgagtaaag...ctacaaataaCTTATCATCGATAAG
  • Template Sequence: atgagtaaaggagaagaacttttca...tggcatggatgagctctacaaataa

pBR322_b View Part

4391nt Extended 4361nt Template

  • Assembly Sequence: gagctctacaaataaCTTATCATCG...TGTTTGACAGgtgagctgataccgc
  • Template Sequence: CTTATCATCGATAAGCTTTAATGCG...CTTCAAGAATTCTCATGTTTGACAG

Primers Download Primers

Below is a summary table of the primers needed for each part of this assembly solution (shown above). Further details on each primer can be viewed by clicking View Primer for each listed below this table.

hp Tm (C): The melting temperature of the primer's hairpin (hp) structure.
hp ΔG: The Gibbs free energy of the primer's hairpin (hp) structure.
hd Tm (C): The melting temperature of the primer's homodimer (hd) structure.
hd ΔG: The Gibbs free energy of the primer's homodimer (hd) structure.
Name Type Tm (C) GC% Hairpin hp Tm (C) hp ΔG Homodimer hd Tm (C) hd ΔG
120714-addgene forward primer fwd 60.471 29.03 True 41.615 -385.626 True 19.277 -7324.651
120714-addgene reverse primer rvs 61.684 65.52 True 35.243 230.700 True 8.420 -5998.441
74625-addgene forward primer fwd 62.012 63.33 True 50.295 -822.084 True 16.903 -7044.691
74625-addgene reverse primer rvs 61.920 50.0 True 39.281 -148.213 True -0.026 -4952.691
Synthetic-1.112 forward primer fwd 59.473 48.39 True 41.613 -271.143 True -3.445 -4572.426
Synthetic-1.112 reverse primer rvs 60.130 34.29 True 53.877 -1047.648 True -23.066 -2929.371
71655-addgene forward primer fwd 60.173 28.95 True 43.634 -565.455 True -24.427 -3386.826
71655-addgene reverse primer rvs 59.620 19.44 True 43.226 -840.354 True 17.553 -7148.451
pBR322_b forward primer fwd 60.806 12.82 True 48.641 -1501.254 True 18.725 -5292.685
pBR322_b reverse primer rvs 60.872 23.08 True 36.900 11.873 True 9.560 -5625.021

Primers List

120714-addgene forward primer View Primer

fwd 60.471°C Hairpin Homodimer

  • Sequence: TCTCATGTTTGACAGgtgagctgataccgct

120714-addgene reverse primer View Primer

rvs 61.684°C Hairpin Homodimer

  • Sequence: gcccttgctcaccatgtcgtgccagctgc

74625-addgene forward primer View Primer

fwd 62.012°C Hairpin Homodimer

  • Sequence: tgcagctggcacgacatggtgagcaagggc

74625-addgene reverse primer View Primer

rvs 61.920°C Hairpin Homodimer

  • Sequence: gatgctggtatcaccttacttgtacagctcgtcc

Synthetic-1.112 forward primer View Primer

fwd 59.473°C Hairpin Homodimer

  • Sequence: gagctgtacaagtaaggtgataccagcatcg

Synthetic-1.112 reverse primer View Primer

rvs 60.130°C Hairpin Homodimer

  • Sequence: ttctcctttactcatattcaacaagaattgggaca

71655-addgene forward primer View Primer

fwd 60.173°C Hairpin Homodimer

  • Sequence: caattcttgttgaatatgagtaaaggagaagaactttt

71655-addgene reverse primer View Primer

rvs 59.620°C Hairpin Homodimer

  • Sequence: CTTATCGATGATAAGttatttgtagagctcatccat

pBR322_b forward primer View Primer

fwd 60.806°C Hairpin Homodimer

  • Sequence: gagctctacaaataaCTTATCATCGATAAGCTTTAATGC

pBR322_b reverse primer View Primer

rvs 60.872°C Hairpin Homodimer

  • Sequence: gcggtatcagctcacCTGTCAAACATGAGAATTCTTGAA
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