scarless test Solution Golden Gate
June 30, 2025, 6:05 p.m.
- 4 Parts 8 Primers
- 60.62C Average Primer Tm 31nt Average Primer 3 BLAST Parts 1 Synthetic Parts 707nt Longest Part 111nt Shortest Part
- 82.00% BLAST nt 18.00% Synthetic nt
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 |
---|---|---|---|---|---|---|---|---|
113412-addgene | addgene | 117 | 136 | 4 | 121 | 14798 | 14666 | 0 |
110282-addgene | addgene | 707 | 741 | 125 | 832 | 9723 | 9013 | 1 |
Synthetic-1.112 | NONE | 111 | 133 | 836 | 947 | 0 | 0 | 2 |
115565-addgene | addgene | 704 | 723 | 951 | 1655 | 8041 | 8757 | 3 |
pBR322_b | NONE | 4360 | 4391 | 1666 | 6026 | 0 | 0 | 4 |
Plasmid Map

Parts List
113412-addgene addgene View Part
136nt Extended 117nt Template
- Assembly Sequence: GGTCTCNGACAGgtgagctgatacc...gcgttggccgattcattaNGAGACC
- Template Sequence: gtgagctgataccgctcgccgcagc...tccccgcgcgttggccgattcatta
110282-addgene addgene View Part
741nt Extended 707nt Template
- Assembly Sequence: GGTCTCNattaatgcagctggcacg...catggacgagctgtacaaNGAGACC
- Template Sequence: atggtgagcaagggcgaggaggata...ccggcggcatggacgagctgtacaa
Synthetic-1.112 View Part
133nt Extended 111nt Template
- Assembly Sequence: GGTCTCNacaagtaaggtgatacca...gtcccaattcttgttgaaNGAGACC
- Template Sequence: ggtgataccagcatcgtcttgatgc...tggagttgtcccaattcttgttgaa
115565-addgene addgene View Part
723nt Extended 704nt Template Internal Cutsite(s) - Adjust probability of success for future experiments
- Assembly Sequence: GGTCTCNtgaatatgagtaaaggag...tacacatggcatggatgaNGAGACC
- Template Sequence: atgagtaaaggagaagaacttttca...ctgggattacacatggcatggatga
pBR322_b View Part
4391nt Extended 4360nt Template Internal Cutsite(s) - Adjust probability of success for future experiments
- Assembly Sequence: GGTCTCNatgaactatacaaataaC...GAATTCTCATGTTTGACANGAGACC
- Template Sequence: CTTATCATCGATAAGCTTTAATGCG...TCTTCAAGAATTCTCATGTTTGACA
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 |
---|---|---|---|---|---|---|---|---|---|
113412-addgene forward primer | fwd | 60.471 | 32.14 | True | 41.615 | -385.626 | True | 19.277 | -7324.651 |
113412-addgene reverse primer | rvs | 60.314 | 33.33 | True | 46.105 | -547.579 | True | 2.079 | -5455.321 |
110282-addgene forward primer | fwd | 62.012 | 45.24 | True | 58.993 | -1443.514 | True | 16.903 | -7044.691 |
110282-addgene reverse primer | rvs | 61.541 | 36.0 | False | 0.000 | 0.000 | True | -8.830 | -3413.286 |
Synthetic-1.112 forward primer | fwd | 59.473 | 35.48 | True | 41.613 | -271.143 | True | 0.084 | -3163.304 |
Synthetic-1.112 reverse primer | rvs | 59.592 | 26.92 | True | 53.877 | -1047.648 | True | -23.073 | -2736.010 |
115565-addgene forward primer | fwd | 60.173 | 22.86 | True | 37.663 | -70.162 | True | -20.930 | -3085.011 |
115565-addgene reverse primer | rvs | 60.961 | 33.33 | False | 0.000 | 0.000 | True | 3.340 | -4081.791 |
pBR322_b forward primer | fwd | 60.806 | 6.25 | True | 48.641 | -1501.254 | True | 18.725 | -5292.685 |
pBR322_b reverse primer | rvs | 60.872 | 0.0 | True | 33.558 | 399.571 | True | 9.560 | -5625.021 |
Primers List
113412-addgene forward primer View Primer
fwd 60.471°C Hairpin Homodimer
- Sequence: GGTCTCNGACAGgtgagctgataccgct
113412-addgene reverse primer View Primer
rvs 60.314°C Hairpin Homodimer
- Sequence: GGTCTCNtaatgaatcggccaacg
110282-addgene forward primer View Primer
fwd 62.012°C Hairpin Homodimer
- Sequence: GGTCTCNattaatgcagctggcacgacatggtgagcaagggc
110282-addgene reverse primer View Primer
rvs 61.541°C Homodimer
- Sequence: GGTCTCNttgtacagctcgtccatg
Synthetic-1.112 forward primer View Primer
fwd 59.473°C Hairpin Homodimer
- Sequence: GGTCTCNacaagtaaggtgataccagcatcg
Synthetic-1.112 reverse primer View Primer
rvs 59.592°C Hairpin Homodimer
- Sequence: GGTCTCNttcaacaagaattgggaca
115565-addgene forward primer View Primer
fwd 60.173°C Hairpin Homodimer
- Sequence: GGTCTCNtgaatatgagtaaaggagaagaactttt
pBR322_b forward primer View Primer
fwd 60.806°C Hairpin Homodimer
- Sequence: GGTCTCNatgaactatacaaataaCTTATCATCGATAAGCTTTAATGC
pBR322_b reverse primer View Primer
rvs 60.872°C Hairpin Homodimer
- Sequence: GGTCTCNTGTCAAACATGAGAATTCTTGAAG