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
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.
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.
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