cs2 test with all db Solution Golden Gate

June 29, 2025, 9:36 p.m.

  • 4 Parts 8 Primers
  • 60.72C Average Primer Tm 30nt Average Primer 3 BLAST Parts 1 Synthetic Parts 717nt Longest Part 112nt Shortest Part
  • 88.00% BLAST nt 12.00% Synthetic nt
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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
115573.1-addgene addgene 133 155 4 137 5252 5120 0
BBa_K2066123-igem igem 711 733 141 852 1949 2659 1
Synthetic-1.112 NONE 112 134 856 968 0 0 2
35346-addgene addgene 717 739 972 1689 1922 2638 3
pBR322_b NONE 4361 4383 1694 6054 0 0 4

Plasmid Map

Parts List

115573.1-addgene addgene View Part

155nt Extended 133nt Template

  • Assembly Sequence: GGTCTCnAGTGgtgagctgataccg...gcagctggcacgacCAGGnGAGACC
  • Template Sequence: gtgagctgataccgctcgccgcagc...gattcattaatgcagctggcacgac

BBa_K2066123-igem igem View Part

733nt Extended 711nt Template

  • Assembly Sequence: GGTCTCnCAGGatggtgagcaaggg...agctgtacaagtaaACTCnGAGACC
  • Template Sequence: atggtgagcaagggcgaagaagata...cggcatggacgagctgtacaagtaa

Synthetic-1.112 View Part

134nt Extended 112nt Template

  • Assembly Sequence: GGTCTCnACTCggtgataccagcat...aattcttgttgaatAAAAnGAGACC
  • Template Sequence: ggtgataccagcatcgtcttgatgc...ggagttgtcccaattcttgttgaat

35346-addgene addgene View Part

739nt Extended 717nt Template

  • Assembly Sequence: GGTCTCnAAAAatgagtaaaggaga...agctctacaaataaAGACnGAGACC
  • Template Sequence: atgagtaaaggagaagaacttttca...tggcatggatgagctctacaaataa

pBR322_b View Part

4383nt Extended 4361nt Template Internal Cutsite(s) - Adjust probability of success for future experiments

  • Assembly Sequence: GGTCTCnAGACCTTATCATCGATAA...CTCATGTTTGACAGAGTGnGAGACC
  • 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
115573.1-addgene forward primer fwd 60.471 33.33 True 48.792 -1260.012 True 16.964 -4571.702
115573.1-addgene reverse primer rvs 61.684 40.0 True 57.817 -2144.785 True 18.461 -5188.669
BBa_K2066123-igem forward primer fwd 62.012 34.62 True 41.611 -254.908 True -24.870 -2693.161
BBa_K2066123-igem reverse primer rvs 61.920 30.0 False 0.000 0.000 True -8.830 -3413.286
Synthetic-1.112 forward primer fwd 59.473 33.33 True 38.109 -59.863 True -30.105 -3125.981
Synthetic-1.112 reverse primer rvs 60.130 22.58 True 53.877 -1047.648 True -23.066 -2929.371
35346-addgene forward primer fwd 60.173 20.59 True 37.663 -70.162 True -20.930 -3085.011
35346-addgene reverse primer rvs 59.620 21.88 False 0.000 0.000 True 17.553 -7148.451
pBR322_b forward primer fwd 60.806 0.0 True 55.376 -2002.494 True 14.497 -5348.232
pBR322_b reverse primer rvs 60.872 0.0 True 33.558 399.571 True 9.560 -5625.021

Primers List

115573.1-addgene forward primer View Primer

fwd 60.471°C Hairpin Homodimer

  • Sequence: GGTCTCnAGTGgtgagctgataccgct

115573.1-addgene reverse primer View Primer

rvs 61.684°C Hairpin Homodimer

  • Sequence: GGTCTCnCCTGgtcgtgccagctgc

BBa_K2066123-igem forward primer View Primer

fwd 62.012°C Hairpin Homodimer

  • Sequence: GGTCTCnCAGGatggtgagcaagggc

BBa_K2066123-igem reverse primer View Primer

rvs 61.920°C Homodimer

  • Sequence: GGTCTCnGAGTttacttgtacagctcgtcc

Synthetic-1.112 forward primer View Primer

fwd 59.473°C Hairpin Homodimer

  • Sequence: GGTCTCnACTCggtgataccagcatcg

Synthetic-1.112 reverse primer View Primer

rvs 60.130°C Hairpin Homodimer

  • Sequence: GGTCTCnTTTTattcaacaagaattgggaca

35346-addgene forward primer View Primer

fwd 60.173°C Hairpin Homodimer

  • Sequence: GGTCTCnAAAAatgagtaaaggagaagaactttt

35346-addgene reverse primer View Primer

rvs 59.620°C Homodimer

  • Sequence: GGTCTCnGTCTttatttgtagagctcatccat

pBR322_b forward primer View Primer

fwd 60.806°C Hairpin Homodimer

  • Sequence: GGTCTCnAGACCTTATCATCGATAAGCTTTAATGC

pBR322_b reverse primer View Primer

rvs 60.872°C Hairpin Homodimer

  • Sequence: GGTCTCnCACTCTGTCAAACATGAGAATTCTTGAA
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