User:Jared Koelling

http://openwetware.org/wiki/840:153g/2008

Part-only sequence for BBa_J45120(1292 bp)

tccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagagtcacacaggaaagtactagatggaagttgttgaagttc ttcacatgaatggaggaaatggagacagtagctatgcaaacaattctttggttcagcaaaaggtgattctcatgacaaagccaataactgagcaagccat gattgatctctacagcagcctctttccagaaaccttatgcattgcagatttgggttgttctttgggagctaacactttcttggtggtctcacagcttgtt aaaatagtagaaaaagaacgaaaaaagcatggttttaagtctccagagttttattttcacttcaatgatcttcctggcaatgattttaatacactttttc agtcactgggggcatttcaagaagatttgagaaagcatataggggaaagctttggtccatgttttttcagtggagtgcctggttcattttatactagact tttcccttccaaaagtttacattttgtttactcctcctacagtctcatgtggctatctcaggtgcctaatgggattgaaaataacaagggaaacatttac atggcaagaacaagccctctaagtgttattaaagcatactacaagcaatatgaaatagatttttcaaattttctcaagtaccgttcagaggaattgatga aaggtggaaagatggtgttaacactcctaggtagagaaagtgaggatcctactagcaaagaatgctgttacatttgggagcttctagccatggccctcaa taagttggttgaagagggattgataaaagaagagaaagtagatgcattcaatattcctcaatacacaccatcaccagcagaagtaaagtacatagttgag aaggaaggatcattcaccattaatcgcttggaaacatcaagagttcattggaatgcttctaataatgagaagaatggtggttacaatgtgtcaaggtgca tgagagctgtggctgagcctttgcttgtcagccactttgacaaggaattgatggatttagtgttccacaagtacgaagagattgtttctgattgcatgtc caaagagaatactgagtttataaatgtcatcatctccttgaccaaaataaattaatactagagccaggcatcaaataaaacgaaaggctcagtcgaaaga ctgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata

Wintergreen – forward and reverse primer #1

Forward Primer Sequence: 	                              ACTACTAGAGTCACACAGGAAAGTA

Primer Start Position: 	53	Primer Length: 	25 Primer TM: 	54.2 ºC 	Primer Self Any: 	6.0 Primer GC %: 	40.0 % 	Primer Self End: 	2.0 Primer End Stability: 	4.30	Primer Penalty: 	16.75

Reverse Primer Sequence: 	                              TCTAGTAGAGAGCGTTCACCGACAA

Primer Start Position: 	1250	Primer Length: 	25 Primer TM: 	58.6 ºC 	Primer Self Any: 	7.0 Primer GC %: 	48.0 % 	Primer Self End: 	2.0 Primer End Stability: 	5.19	Primer Penalty: 	4.39

Forward primer 1 will have restriction site Xba1 added (TCTAGA)with 4 random nucleotides added for the sequence of AACATCTAGA Reverse primer 1 will have restriction site Spe1 added (ACTAGT) with 4 random nucleotides added for the sequence of TAATACTAGT

For a total forward primer 1 of AACATCTAGAACTACTAGAGTCACACAGGAAAGTA For a total reverse primer 1 of TAATACTAGTTCTAGTAGAGAGCGTTCACCGACAA

tccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagagtcacacaggaaagtactagatggaagttgttgaagttc ttcacatgaatggaggaaatggagacagtagctatgcaaacaattctttggttcagcaaaaggtgattctcatgacaaagccaataactgagcaagccat gattgatctctacagcagcctctttccagaaaccttatgcattgcagatttgggttgttctttgggagctaacactttcttggtggtctcacagcttgtt aaaatagtagaaaaagaacgaaaaaagcatggttttaagtctccagagttttattttcacttcaatgatcttcctggcaatgattttaatacactttttc agtcactgggggcatttcaagaagatttgagaaagcatataggggaaagctttggtccatgttttttcagtggagtgcctggttcattttatactagact tttcccttccaaaagtttacattttgtttactcctcctacagtctcatgtggctatctcaggtgcctaatgggattgaaaataacaagggaaacatttac atggcaagaacaagccctctaagtgttattaaagcatactacaagcaatatgaaatagatttttcaaattttctcaagtaccgttcagaggaattgatga aaggtggaaagatggtgttaacactcctaggtagagaaagtgaggatcctactagcaaagaatgctgttacatttgggagcttctagccatggccctcaa taagttggttgaagagggattgataaaagaagagaaagtagatgcattcaatattcctcaatacacaccatcaccagcagaagtaaagtacatagttgag aaggaaggatcattcaccattaatcgcttggaaacatcaagagttcattggaatgcttctaataatgagaagaatggtggttacaatgtgtcaaggtgca tgagagctgtggctgagcctttgcttgtcagccactttgacaaggaattgatggatttagtgttccacaagtacgaagagattgtttctgattgcatgtc caaagagaatactgagtttataaatgtcatcatctccttgaccaaaataaattaatactagagccaggcatcaaataaaacgaaaggctcagtcgaaaga ctgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata

Wintergreen -- forward and reverse primer #2 Forward Primer Sequence: 	                                  TGAGCACTACTAGAGTCACACAGG Primer Start Position: 	48	Primer Length: 	24 Primer TM: 	57.3 ºC 	Primer Self Any: 	7.0 Primer GC %: 	50.0 % 	Primer Self End: 	1.0 Primer End Stability: 	6.01	Primer Penalty: 	2.69

Reverse Primer Sequence: 	                                  AGTAGAGAGCGTTCACCGACAAAC Primer Start Position: 	1247	Primer Length: 	24 Primer TM: 	58.5 ºC 	Primer Self Any: 	4.0 Primer GC %: 	50.0 % 	Primer Self End: 	2.0 Primer End Stability: 	4.89	Primer Penalty: 	1.53 Forward primer 2 will have restriction site Xba1 added (TCTAGA) with 4 random nucleotides added for the sequence of TAATTCTAGA Reverse primer 2 will have restriction site Pst1 added (CTGCAG) with 4 random nucleotides added for the sequence of TAATCTGCAG

For a total forward primer 2 of TAATTCTAGATGAGCACTACTAGAGTCACACAGG For a total reverse primer 2 of TAATCTGCAGAGTAGAGAGCGTTCACCGACAAAC