LOCUS       pSEM270___MCS_mo        3150 bp    DNA        circular     14-AUG-2020
DEFINITION  .
COMMENT     Unpublished reagents. Sonia el Mouridi and Christian
            Froekjaer-Jensen. Synthetic Genome Biology lab. KAUST.
COMMENT     www.wormbuilder.org
COMMENT     MOdular Safe harbor Transgene Insertion (mosTI). 
COMMENT     Multiple cloning site and Golden Gate compatible (BsaI) vector for
            use with split hygroR selection.
COMMENT     Use for single-copy transgene insertion at landing site.
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     primer_bind     48..69
                     /locus_tag="UniF"
                     /label="UniF"
                     /ApEinfo_label="UniF"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1..17
                     /locus_tag="M13F"
                     /label="M13F"
                     /ApEinfo_label="M13F"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(1055..1071)
                     /locus_tag="M13R"
                     /label="M13R"
                     /ApEinfo_label="M13R"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(999..1019)
                     /locus_tag="UniR"
                     /label="UniR"
                     /ApEinfo_label="UniR"
                     /ApEinfo_fwdcolor="#72fa78"
                     /ApEinfo_revcolor="#ff7d78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    2072..3037
                     /locus_tag="AmpR"
                     /label="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#929292"
                     /ApEinfo_revcolor="#929292"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1243..1831
                     /locus_tag="Ori"
                     /label="Ori"
                     /ApEinfo_label="Ori"
                     /ApEinfo_fwdcolor="#d5d5d5"
                     /ApEinfo_revcolor="#d5d5d5"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    699..998
                     /locus_tag="Spacer (no stop, no PolyA)"
                     /label="Spacer (no stop, no PolyA)"
                     /ApEinfo_label="Spacer (no stop, no PolyA)"
                     /ApEinfo_fwdcolor="#75d5ff"
                     /ApEinfo_revcolor="#75d5ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    574..607
                     /locus_tag="loxP"
                     /label="loxP"
                     /ApEinfo_label="loxP"
                     /ApEinfo_fwdcolor="#ffd478"
                     /ApEinfo_revcolor="#fefc78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     3'UTR           533..573
                     /locus_tag="rps-12 3' UTR"
                     /label="rps-12 3' UTR"
                     /ApEinfo_label="rps-12 3' UTR"
                     /ApEinfo_fwdcolor="#f76ca1"
                     /ApEinfo_revcolor="#d783ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             join(70..285,442..532)
                     /locus_tag="HygroR (non-rescuing fragment)"
                     /label="HygroR (non-rescuing fragment)"
                     /ApEinfo_label="HygroR (non-rescuing fragment)"
                     /ApEinfo_fwdcolor="#fefc78"
                     /ApEinfo_revcolor="#fefc78"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    613..683
                     /locus_tag="MCS(w 2xBsaI)"
                     /label="MCS(w 2xBsaI)"
                     /ApEinfo_label="MCS(w 2xBsaI)"
                     /ApEinfo_fwdcolor="#28ad48"
                     /ApEinfo_revcolor="#28ad48"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
ORIGIN
        1 gtaaaacgac ggccagtcat gcataatccg cacgcatctg gaataaggaa gtgccattcc
       61 gcctgacctA TGCATGGAAC AACAAACCAG ATATTTTGAA CGACGACACC CAGAATTAGC
      121 CGGATCTCCT AGGCTTCGAG CTTACATGCT TAGAATCGGA TTAGATCAGT TGTACCAATC
      181 TCTTGTGGAT GGTAACTTTG ACGACGCTGC GTGGGCTCAA GGGCGTTGTG ATGCCATTGT
      241 GCGTTCAGGG GCAGGCACGG TAGGACGAAC TCAGATTGCA AGAAGgtaag ttttcaaaaa
      301 attaagattc agttatggga agtgggagct tacatatttt caggctttcc ctatccaaaa
      361 cgctcgttcg agtgtttcag ttcggtatgt cgaggcgtta ccggacggaa aatatttaaa
      421 ataaaatcct cgtcatttca gGTCAGCTGC GGTTTGGACT GACGGATGCG TCGAGGTCCT
      481 TGCTGACTCA GGTAATAGAA GACCTTCTAC AAGACCACGT GCCAAAGAGT AAtctgttcg
      541 ttgttccaat aaattgttat tgttccctat ttgataactt cgtatagcat acattatacg
      601 aagttattgc ctGAGACCAG CTAGCAAGCT TGCATGCCTG CAGGTCGACT CTAGAGGATC
      661 CCCGGGTACC GAGCTCAGGT CTCtgagctg gcgggagctc aggaaacgga gaatctgtgc
      721 ggcaatgtct tatcatttga aacgcgccgt atcgatgctg acatcatgcg gctcccgtgt
      781 tagtcgagaa gatgggtacg ggcctttatt catccaacac tctacggctt cttcaagagc
      841 tagtagggca ccctgtagtt ggaaagggat tatttcgtat ggcgagccca taccgtcttt
      901 cttgcggaag cttaacacga taggaagttg gatagtttcg aacgatggtt ccatacggaa
      961 cgctgtctca aggatgagtc tgacggagtg taactcgaag gctaggtgga ggctcagtga
     1021 tgataagtct gcgatggtgg atgcatgtgt catggtcata gctgtttcct gtgtgaaatt
     1081 gttatccgct cagagggcac aatcctattc cgcgctatcc gacaatctcc aagacattag
     1141 gtggagttca gttcggcgta tggcatatgt cgctggaaag aacatgtgag caaaaggcca
     1201 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1261 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1321 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1381 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1441 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1501 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1561 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1621 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1681 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1861 tgacgctcta ttcaacaaag ccgccgtccc gtcaagtcag cgtaaatggg tagggggctt
     1921 caaatcgtcc tcgtgatacc aattcggagc ctgctttttt gtacaaactt gttgataatg
     1981 gcaattcaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2041 aaagtatata tgagtaaact tggtctgaca gTTACCAATG CTTAATCAGT GAGGCACCTA
     2101 TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG ACTCCCCGTC GTGTAGATAA
     2161 CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC AATGATACCG CGAGAGCCAC
     2221 GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC CGGAAGGGCC GAGCGCAGAA
     2281 GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA TTGTTGCCGG GAAGCTAGAG
     2341 TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC CATTGCTACA GGCATCGTGG
     2401 TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG TTCCCAACGA TCAAGGCGAG
     2461 TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC CTTCGGTCCT CCGATCGTTG
     2521 TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT GGCAGCACTG CATAATTCTC
     2581 TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG TGAGTACTCA ACCAAGTCAT
     2641 TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC GGCGTCAATA CGGGATAATA
     2701 CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG AAAACGTTCT TCGGGGCGAA
     2761 AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT GTAACCCACT CGTGCACCCA
     2821 ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG GTGAGCAAAA ACAGGAAGGC
     2881 AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG TTGAATACTC ATACTCTTCC
     2941 TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT CATGAGCGGA TACATATTTG
     3001 AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGcac atttccccga aaagtgccag
     3061 atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt gatccaccac
     3121 aagcgccagg gttttcccag tcacgacgtt   
//
