CS2 25nt-tail Solution PCR

July 24, 2025, 5:31 p.m.

  • 4 Parts 8 Primers
  • 60.88C Average Primer Tm 31nt Average Primer 3 BLAST Parts 1 Synthetic Parts 717nt Longest Part 112nt Shortest Part
  • 80.00% BLAST nt 20.00% 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
113429-addgene addgene 133 157 0 133 16833 16701 0
75370-addgene addgene 711 735 133 844 8331 9041 1
Synthetic-1.112 NONE 112 136 844 956 0 0 2
79550-addgene addgene 717 741 956 1673 1132 1848 3
pBR322_b NONE 4361 4385 1674 6034 0 0 4

Plasmid Map

Parts List

113429-addgene addgene View Part

157nt Extended 133nt Template

  • Assembly Sequence: CATGTTTGACAGgtgagctgatacc...cagctggcacgacatggtgagcaag
  • Template Sequence: gtgagctgataccgctcgccgcagc...gattcattaatgcagctggcacgac

75370-addgene addgene View Part

735nt Extended 711nt Template

  • Assembly Sequence: agctggcacgacatggtgagcaagg...gctgtacaagtaaggtgataccagc
  • Template Sequence: atggtgagcaagggcgaggaggata...cggcatggacgagctgtacaagtaa

Synthetic-1.112 View Part

136nt Extended 112nt Template

  • Assembly Sequence: ctgtacaagtaaggtgataccagca...attcttgttgaatatgagtaaagga
  • Template Sequence: ggtgataccagcatcgtcttgatgc...ggagttgtcccaattcttgttgaat

79550-addgene addgene View Part

741nt Extended 717nt Template

  • Assembly Sequence: ttcttgttgaatatgagtaaaggag...tctctacaaataaCTTATCATCGAT
  • Template Sequence: atgagtaaaggagaagaacttttca...tggcatggatgatctctacaaataa

pBR322_b View Part

4385nt Extended 4361nt Template

  • Assembly Sequence: ctctacaaataaCTTATCATCGATA...TCATGTTTGACAGgtgagctgatac
  • 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
113429-addgene forward primer fwd 60.471 32.14 True 41.615 -385.626 True 19.277 -7324.651
113429-addgene reverse primer rvs 61.684 61.54 True 35.243 230.700 True 8.420 -5998.441
75370-addgene forward primer fwd 62.012 62.96 True 38.757 -227.570 True 13.766 -2486.263
75370-addgene reverse primer rvs 61.920 51.61 True 39.281 -148.213 True -7.305 -4332.761
Synthetic-1.112 forward primer fwd 59.473 46.43 True 41.613 -271.143 True -6.869 -3703.136
Synthetic-1.112 reverse primer rvs 60.130 34.38 True 53.877 -1047.648 True -23.066 -2929.371
79550-addgene forward primer fwd 60.173 28.57 True 37.663 -70.162 True -20.930 -3085.011
79550-addgene reverse primer rvs 61.240 22.86 True 41.443 -613.005 True -8.509 -4337.151
pBR322_b forward primer fwd 60.806 8.33 True 48.641 -1501.254 True 18.725 -5292.685
pBR322_b reverse primer rvs 60.872 16.67 True 36.900 11.873 True 9.560 -5625.021

Primers List

113429-addgene forward primer View Primer

fwd 60.471°C Hairpin Homodimer

  • Sequence: CATGTTTGACAGgtgagctgataccgct

113429-addgene reverse primer View Primer

rvs 61.684°C Hairpin Homodimer

  • Sequence: cttgctcaccatgtcgtgccagctgc

75370-addgene forward primer View Primer

fwd 62.012°C Hairpin Homodimer

  • Sequence: agctggcacgacatggtgagcaagggc

75370-addgene reverse primer View Primer

rvs 61.920°C Hairpin Homodimer

  • Sequence: gctggtatcaccttacttgtacagctcgtcc

Synthetic-1.112 forward primer View Primer

fwd 59.473°C Hairpin Homodimer

  • Sequence: ctgtacaagtaaggtgataccagcatcg

Synthetic-1.112 reverse primer View Primer

rvs 60.130°C Hairpin Homodimer

  • Sequence: tcctttactcatattcaacaagaattgggaca

79550-addgene forward primer View Primer

fwd 60.173°C Hairpin Homodimer

  • Sequence: ttcttgttgaatatgagtaaaggagaagaactttt

79550-addgene reverse primer View Primer

rvs 61.240°C Hairpin Homodimer

  • Sequence: ATCGATGATAAGttatttgtagagatcatccatgc

pBR322_b forward primer View Primer

fwd 60.806°C Hairpin Homodimer

  • Sequence: ctctacaaataaCTTATCATCGATAAGCTTTAATGC

pBR322_b reverse primer View Primer

rvs 60.872°C Hairpin Homodimer

  • Sequence: gtatcagctcacCTGTCAAACATGAGAATTCTTGAA
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