scarless test Solution Golden Gate
June 30, 2025, 6:05 p.m.
- 4 Parts 8 Primers
- 60.42C Average Primer Tm 34nt Average Primer 3 BLAST Parts 1 Synthetic Parts 698nt Longest Part 111nt Shortest Part
- 90.00% BLAST nt 10.00% 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 |
---|---|---|---|---|---|---|---|---|
116061-addgene | addgene | 117 | 156 | 4 | 121 | 7199 | 7067 | 0 |
64128-addgene | addgene | 698 | 732 | 125 | 823 | 5036 | 5746 | 1 |
Synthetic-1.112 | NONE | 111 | 142 | 827 | 938 | 0 | 0 | 2 |
35363-addgene | addgene | 693 | 712 | 942 | 1635 | 5726 | 6442 | 3 |
pBR322_b | NONE | 4340 | 4382 | 1675 | 6035 | 0 | 0 | 4 |
Plasmid Map

Parts List
116061-addgene addgene View Part
156nt Extended 117nt Template
- Assembly Sequence: GGTCTCNCTTCAAGAATTCTCATGT...gcgttggccgattcattaNGAGACC
- Template Sequence: gtgagctgataccgctcgccgcagc...tccccgcgcgttggccgattcatta
64128-addgene addgene View Part
732nt Extended 698nt Template
- Assembly Sequence: GGTCTCNattaatgcagctggcacg...caccggcggcatggacgaNGAGACC
- Template Sequence: atggtgagcaagggcgaggaggata...gccactccaccggcggcatggacga
Synthetic-1.112 View Part
142nt Extended 111nt Template
- Assembly Sequence: GGTCTCNacgagctgtacaagtaag...gtcccaattcttgttgaaNGAGACC
- Template Sequence: ggtgataccagcatcgtcttgatgc...tggagttgtcccaattcttgttgaa
35363-addgene addgene View Part
712nt Extended 693nt Template
- Assembly Sequence: GGTCTCNtgaatatgagtaaaggag...gctgctgggattacacatNGAGACC
- Template Sequence: atgagtaaaggagaagaacttttca...tgtaactgctgctgggattacacat
pBR322_b View Part
4382nt Extended 4340nt Template Internal Cutsite(s) - Adjust probability of success for future experiments
- Assembly Sequence: GGTCTCNacatggcatggatgagct...CGAGGCCCTTTCGTCTTCNGAGACC
- Template Sequence: CTTATCATCGATAAGCTTTAATGCG...CGTATCACGAGGCCCTTTCGTCTTC
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 |
---|---|---|---|---|---|---|---|---|---|
116061-addgene forward primer | fwd | 60.471 | 18.75 | True | 41.615 | -385.626 | True | 19.277 | -7324.651 |
116061-addgene reverse primer | rvs | 60.314 | 33.33 | True | 46.105 | -547.579 | True | 2.079 | -5455.321 |
64128-addgene forward primer | fwd | 62.012 | 45.24 | True | 58.993 | -1443.514 | True | 16.903 | -7044.691 |
64128-addgene reverse primer | rvs | 60.583 | 45.0 | False | 0.000 | 0.000 | True | -27.504 | -2497.791 |
Synthetic-1.112 forward primer | fwd | 59.473 | 40.0 | True | 37.584 | -92.277 | True | 18.599 | -1553.373 |
Synthetic-1.112 reverse primer | rvs | 59.592 | 26.92 | True | 53.877 | -1047.648 | True | -23.073 | -2736.010 |
35363-addgene forward primer | fwd | 60.173 | 22.86 | True | 37.663 | -70.162 | True | -20.930 | -3085.011 |
35363-addgene reverse primer | rvs | 59.208 | 33.33 | False | 0.000 | 0.000 | True | -33.069 | -2810.310 |
pBR322_b forward primer | fwd | 60.806 | 18.64 | True | 42.695 | -1074.653 | True | 29.241 | -9147.601 |
pBR322_b reverse primer | rvs | 61.570 | 0.0 | True | 50.507 | -1289.519 | True | -4.088 | -5260.081 |
Primers List
116061-addgene forward primer View Primer
fwd 60.471°C Hairpin Homodimer
- Sequence: GGTCTCNCTTCAAGAATTCTCATGTTTGACAGgtgagctgataccgct
116061-addgene reverse primer View Primer
rvs 60.314°C Hairpin Homodimer
- Sequence: GGTCTCNtaatgaatcggccaacg
64128-addgene forward primer View Primer
fwd 62.012°C Hairpin Homodimer
- Sequence: GGTCTCNattaatgcagctggcacgacatggtgagcaagggc
Synthetic-1.112 forward primer View Primer
fwd 59.473°C Hairpin Homodimer
- Sequence: GGTCTCNacgagctgtacaagtaaggtgataccagcatcg
Synthetic-1.112 reverse primer View Primer
rvs 59.592°C Hairpin Homodimer
- Sequence: GGTCTCNttcaacaagaattgggaca
35363-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: GGTCTCNacatggcatggatgagctctacaaataaCTTATCATCGATAAGCTTTAATGC
pBR322_b reverse primer View Primer
rvs 61.570°C Hairpin Homodimer
- Sequence: GGTCTCNGAAGACGAAAGGGCCT