LOCUS       Complete_construct_plasmid_map_-_P2d        6688 bp    DNA     circular UNA 05-SEP-2022
DEFINITION  Gibson/Homology Cloning ligation product.
ACCESSION   urn.local.elinlarsson.2k-f9qmcqg
VERSION     urn.local.elinlarsson.2k-f9qmcqg
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    382..580
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="right end of Tn7"
     misc_feature    629..723
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
                     /label="bacteriophage lambda T0 terminator"
     misc_feature    820..917
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
                     /label="Escherichia coli rrnB T1 terminator"
     misc_feature    958..1005
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="FRT"
     CDS             complement(1135..1668)
                     /ApEinfo_revcolor="#4444ff"
                     /ApEinfo_fwdcolor="#4444ff"
                     /label="aacC1"
     CDS             complement(1135..1668)
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /label="GmR"
     misc_feature    1926..1973
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="FRT"
     primer_bind     2010..2030
                     /Sequence="TCAAAGAGTGGAACAATGCAG"
                     /Tm="56.7"
                     /%GC="42.9"
                     /Hairpin_Tm="44.2"
                     /Self_Dimer_Tm="None"
                     /created_by="elinlarsson"
                     /note="Derived using Geneious Prime 2022.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pJM220-mNeongreen not codon optimized
                     backbone piece - NZ_CP027755.1 - Annotated P. synxantha
                     2-79 from pseudomonas.com (reversed) PCR Product"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; col_F
                     (<a href=""urn:local:.:ti-em03ja7"">pJM220-mNeongreen not
                     codon optimized backbone piece - NZ_CP027755.1 - Annotated
                     P. synxantha 2-79 from pseudomonas.com (reversed) PCR
                     Product</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     col_F (<a
                     href=""urn:local:.:vm-em1636y"">pJM220-mNeongreen not
                     codon optimized backbone piece - NZ_CP027755.1 - Annotated
                     P. synxantha 2-79 from pseudomonas.com extraction
                     (reversed) PCR Product</a>)"
                     /label="col_F"
     primer_bind     complement(2075..2094)
                     /Sequence="ACGATGTTAGCGGCGACAAT"
                     /Tm="60.5"
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="13.0"
                     /created_by="elinlarsson"
                     /note="Derived using Geneious Prime 2022.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pJM220-T0T1::PrhaBAD-mN extraction"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     EL_pJM220NG_bb_R (<a
                     href=""urn:local:.:bi-eoqp3mw"">pJM220-T0T1::PrhaBAD-mN
                     extraction</a>)"
                     /label="EL_pJM220NG_bb_R"
     primer_bind     complement(2084..2102)
                     /Sequence="CGATGCCGACGATGTTAGC"
                     /Tm="59.2"
                     /%GC="57.9"
                     /Hairpin_Tm="38.2"
                     /Self_Dimer_Tm="None"
                     /created_by="elinlarsson"
                     /note="Derived using Geneious Prime 2022.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pJM220-T0T1::PrhaBAD-mS extraction - 1.
                     pJM220-T0T1::pPhoA_stro extraction - 2c.
                     pJM220-T0T1::PrhaBAD-mN extraction - mNeonGreen - 3.
                     miniTn7(Gm)PA1/04/03-ecfp - lambda t0 terminator - 4.
                     miniTn7(Gm)PA1/04/03-ecfp - Pa1/04/03"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     pJM220_mS_R (<a
                     href=""urn:local:.:dw-eoqpkff"">pJM220-T0T1::PrhaBAD-mS
                     extraction - 1. pJM220-T0T1::pPhoA_stro extraction - 2c.
                     pJM220-T0T1::PrhaBAD-mN extraction - mNeonGreen - 3.
                     miniTn7(Gm)PA1/04/03-ecfp - lambda t0 terminator - 4.
                     miniTn7(Gm)PA1/04/03-ecfp - Pa1/04/03</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:fb-eor906y"">pJM220_mS_R</a>"
                     /label="pJM220_mS_R"
     ligation        2085..2102
                     /label="Ligation"
     misc_feature    2085..2249
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:1t2-f5bxdli"
                     /label="1. pJM220-T0T1::pPhoA_stro extraction
                     (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     2103..2125
                     /annotation_group="284313174897451e9e0a16b-d6e9-4774-8463-
                     43c6298dab38"
                     /Sequence="AGAGCGCTTTTGAAGCTAATTCG"
                     /created_by="primer3"
                     /Product_Size=138
                     /#_Off-target_Sites=0
                     /Off-target_Template="1. pJM220-T0T1::pPhoA_stro
                     extraction"
                     /Extension="CTAACATCGTCGGCATCG"
                     /Tm_(binding_region)="60.2"
                     /Tm_(with_extension)="67.0"
                     /%GC_(binding_region)="43.5"
                     /%GC_(with_extension)="48.8"
                     /Hairpin_Tm_(with_extension)="43.8"
                     /Self_Dimer_Tm_(with_extension)="26.4"
                     /Pair_Dimer_Tm_(binding_region)="None"
                     /label="1. pJM220-T0T1::pPhoA_stro extraction (1 F) -
                     pJM220-T0T1::PrhaBAD-mS extraction PCR Product"
     CDS             2133..2216
                     /created_by="elinlarsson"
                     /note="Derived using Geneious Prime 2022.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PphoA_K1139200 (iGem)"
                     /Transferred_Similarity="100.00%"
                     /Adjusted_Transferred_Similarity="83.33%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:9/6 (adjusted
                     to 83.33%; Adjusted-Interval-0:0/0); iGem phoA promoter
                     (<a href=""urn:local:.:bw-e0s7nai"">PphoA_K1139200
                     (iGem)</a>)"
                     /label="iGem phoA promoter"
     source          2142..2222
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /label="Ecoli pPhoA"
     primer_bind     complement(2196..2240)
                     /Sequence="GCTTAATTTCTCCTCTTTAATACATTAAAAAATAAAAACAAAGCG"
                     /Tm="58.0"
                     /%GC="24.4"
                     /Hairpin_Tm="35.7"
                     /Self_Dimer_Tm="3.8"
                     /created_by="elinlarsson"
                     /label="phoA_bb_R"
     primer_bind     complement(2217..2240)
                     /annotation_group="284313528168011e8d6dc3f-4e4f-4700-8964-
                     522f6ee6435b"
                     /Sequence="GCTTAATTTCTCCTCTTTAATACA"
                     /created_by="primer3"
                     /Product_Size=138
                     /#_Off-target_Sites=0
                     /Off-target_Template="1. pJM220-T0T1::pPhoA_stro
                     extraction"
                     /Extension="GGAGACCAT"
                     /Tm_(binding_region)="52.9"
                     /Tm_(with_extension)="62.5"
                     /%GC_(binding_region)="29.2"
                     /%GC_(with_extension)="36.4"
                     /Hairpin_Tm_(with_extension)="None"
                     /Self_Dimer_Tm_(with_extension)="None"
                     /Pair_Dimer_Tm_(binding_region)="None"
                     /label="1. pJM220-T0T1::pPhoA_stro extraction (138 R) -
                     2c. pJM220-T0T1::PrhaBAD-mN extraction - mNeonGreen"
     RBS             2223..2234
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /label="strong bacterial ribosome binding site (Elowitz
                     and Leibler, 2000)"
     misc_feature    2223..2240
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /label="PA1/04/03 fragment"
     ligation        2232..2249
                     /label="Ligation"
     misc_feature    2232..2961
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:2q-f9qmcql"
                     /label="2c. pJM220-T0T1::PrhaBAD-mN extraction -
                     mNeonGreen (concatenated sequence 3)"
                     /note="Geneious type: Concatenated sequence"
     primer          <2241..2254
                     /note="sequence:
                     actagtgctctgcagtaaggaggaacagctATGGTCTCCAAGGG"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="oGS32"
     CDS             2241..2951
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /label="mNeonGreen"
     primer_bind     2241..2259
                     /annotation_group="284313815745136ad9f096a-ffa8-4c90-8e26-
                     b8ec0f787e87"
                     /Sequence="ATGGTCTCCAAGGGCGAGG"
                     /created_by="primer3"
                     /Product_Size=711
                     /#_Off-target_Sites=0
                     /Off-target_Template="2c. pJM220-T0T1::PrhaBAD-mN
                     extraction - mNeonGreen"
                     /Extension="AAATTAAGC"
                     /Tm_(binding_region)="61.4"
                     /Tm_(with_extension)="67.1"
                     /%GC_(binding_region)="63.2"
                     /%GC_(with_extension)="50.0"
                     /Hairpin_Tm_(with_extension)="37.0"
                     /Self_Dimer_Tm_(with_extension)="None"
                     /Pair_Dimer_Tm_(binding_region)="2.4"
                     /label="2c. pJM220-T0T1::PrhaBAD-mN extraction -
                     mNeonGreen (1 F) - 1. pJM220-T0T1::pPhoA_stro extraction"
     primer_bind     2241..2258
                     /Sequence="ATGGTCTCCAAGGGCGAG"
                     /Tm="58.7"
                     /%GC="61.1"
                     /Hairpin_Tm="37.0"
                     /Self_Dimer_Tm="None"
                     /created_by="elinlarsson"
                     /label="mNG_F"
     misc_feature    2921..3149
                     /created_by="elinlarsson"
                     /label="fEL_010_gBlock"
     primer_bind     complement(2926..2951)
                     /created_by="elinlarsson"
                     /Sequence="TTACTTATAGAGCTCATCCATCCCCA"
                     /Tm="60.7"
                     /%GC="42.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="17.8"
                     /modified_by="elinlarsson"
                     /label="mNG_R"
     primer_bind     complement(2927..2951)
                     /annotation_group="284314139737530651abdf4-c358-4e21-aa0a-
                     3c0d5bc3f58f"
                     /Sequence="TTACTTATAGAGCTCATCCATCCCC"
                     /created_by="primer3"
                     /Product_Size=711
                     /#_Off-target_Sites=0
                     /Off-target_Template="2c. pJM220-T0T1::PrhaBAD-mN
                     extraction - mNeonGreen"
                     /Extension="AACAGGAGTC"
                     /Tm_(binding_region)="59.3"
                     /Tm_(with_extension)="68.8"
                     /%GC_(binding_region)="44.0"
                     /%GC_(with_extension)="45.7"
                     /Hairpin_Tm_(with_extension)="None"
                     /Self_Dimer_Tm_(with_extension)="16.2"
                     /Pair_Dimer_Tm_(binding_region)="2.4"
                     /label="2c. pJM220-T0T1::PrhaBAD-mN extraction -
                     mNeonGreen (711 R) - 3. miniTn7(Gm)PA1/04/03-ecfp - lambda
                     t0 terminator"
     primer_bind     complement(2927..2948)
                     /Sequence="CTTATAGAGCTCATCCATCCCC"
                     /Tm="57.2"
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="17.8"
                     /created_by="elinlarsson"
                     /label="bb_R"
     primer          complement(2932..>2951)
                     /note="sequence:
                     atcagaccgcttctgcgttctgattcgcgaTTACTTATAGAGCTCATCCA"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="oGS33"
     ligation        2942..2961
                     /label="Ligation"
     misc_feature    2942..>3046
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:2r-f9qmcqm"
                     /label="3. miniTn7(Gm)PA1/04/03-ecfp - lambda t0
                     terminator (concatenated sequence 4)"
                     /note="Geneious type: Concatenated sequence"
     source          <2952..>3046
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /label="source"
     terminator      2952..3046
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /label="lambda t0 terminator"
     primer_bind     2952..2979
                     /created_by="elinlarsson"
                     /Sequence="GACTCCTGTTGATAGATCCAGTAATGAC"
                     /Tm="60.9"
                     /%GC="42.9"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /modified_by="elinlarsson"
                     /label="term_P2d_F"
     primer_bind     2952..2973
                     /annotation_group="284314869391325e0f3feaa-2ae7-4122-9f30-
                     2b03697e58c1"
                     /Sequence="GACTCCTGTTGATAGATCCAGT"
                     /created_by="primer3"
                     /Product_Size=95
                     /#_Off-target_Sites=0
                     /Off-target_Template="3. miniTn7(Gm)PA1/04/03-ecfp -
                     lambda t0 terminator"
                     /Extension="CTATAAGTAA"
                     /Tm_(binding_region)="56.7"
                     /Tm_(with_extension)="61.7"
                     /%GC_(binding_region)="45.5"
                     /%GC_(with_extension)="37.5"
                     /Hairpin_Tm_(with_extension)="None"
                     /Self_Dimer_Tm_(with_extension)="None"
                     /Pair_Dimer_Tm_(binding_region)="None"
                     /label="3. miniTn7(Gm)PA1/04/03-ecfp - lambda t0
                     terminator (1 F) - 2c. pJM220-T0T1::PrhaBAD-mN extraction
                     - mNeonGreen"
     primer_bind     complement(3023..3046)
                     /annotation_group="284315157465747d3612d66-736b-422c-9a49-
                     b2a47fb436dd"
                     /Sequence="ATTCTCACCAATAAAAAACGCCCG"
                     /created_by="primer3"
                     /Product_Size=95
                     /#_Off-target_Sites=0
                     /Off-target_Template="3. miniTn7(Gm)PA1/04/03-ecfp -
                     lambda t0 terminator"
                     /Extension="TTTTGATAAA"
                     /Tm_(binding_region)="60.6"
                     /Tm_(with_extension)="65.0"
                     /%GC_(binding_region)="41.7"
                     /%GC_(with_extension)="32.4"
                     /Hairpin_Tm_(with_extension)="44.8"
                     /Self_Dimer_Tm_(with_extension)="None"
                     /Pair_Dimer_Tm_(binding_region)="None"
                     /label="3. miniTn7(Gm)PA1/04/03-ecfp - lambda t0
                     terminator (95 R) - 4. miniTn7(Gm)PA1/04/03-ecfp -
                     Pa1/04/03"
     ligation        3037..>3046
                     /label="Ligation"
     misc_feature    3037..3143
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:1sk-f5bxdli"
                     /label="4. miniTn7(Gm)PA1/04/03-ecfp - Pa1/04/03
                     (concatenated sequence 5)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    3047..3124
                     /Original_Bases="TTTATCAAAAAGAGTGTTGACTTGTGAGCGGATAACAATGA
                     TACTTAGATTCAATTGTGAGCGGATAACAATTTCACACA"
                     /label="TTTATCAAAAAGAGTGTTGACTTGTGAGCGGATAACAATGATACTTAGAT
                     TCAATTGTGAGCGGATAACAATTTCACACA"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    <3047..>3124
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:1ws-f5bxdll"
                     /label="P2d PCR Product (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    join(3106..>3124,<3047..3062)
                     /ApEinfo_revcolor="#6ecff6"
                     /ApEinfo_fwdcolor="#6ecff6"
                     /label="DVLA"
     misc_feature    join(3107..>3124,<3047..3061)
                     /Color="#cc99ff"
                     /Transferred_From="CIDAR_DVA_backbone"
                     /Transferred_Similarity="100.00%"
                     /label="CIDAR DVA backbone"
     primer_bind     3047..3067
                     /annotation_group="3366786197007798247e72f-047a-44df-b07f-
                     e74c1c7cca4c"
                     /Sequence="AGACTAGTGGGTCTCAGGAGT"
                     /created_by="primer3"
                     /Product_Size=78
                     /#_Off-target_Sites=0
                     /Off-target_Template="P2d PCR Product"
                     /Extension="TGTCGCCGCTAACATCGT"
                     /Tm_(binding_region)="59.0"
                     /Tm_(with_extension)="68.2"
                     /%GC_(binding_region)="52.4"
                     /%GC_(with_extension)="53.8"
                     /Hairpin_Tm_(with_extension)="40.2"
                     /Self_Dimer_Tm_(with_extension)="None"
                     /Pair_Dimer_Tm_(binding_region)="None"
                     /label="P2d PCR Product (1 F) - pJM220-mNeongreen not
                     codon optimized backbone piece"
     promoter        3067..3101
                     /Color="#ffffff"
                     /modified_by="murraylab"
                     /created_by="Andrey"
                     /Transferred_From="CIDAR_weak_promoter_J23116, J23116
                     Promoter (0.16x)"
                     /Transferred_Similarity="100.00%"
                     /label="CIDAR weak promoter J23116 (396x); J23116 Promoter
                     (0.16x) "
     primer_bind     complement(3103..3124)
                     /annotation_group="33667895288358804ffb5f5-eee4-4b90-ad51-
                     5d9dc0b31199"
                     /Sequence="CCGCTACTAGTAGGTCTCTAGT"
                     /created_by="primer3"
                     /Product_Size=78
                     /#_Off-target_Sites=0
                     /Off-target_Template="P2d PCR Product"
                     /Extension="TGCTTAATTTCTCCTCTTT"
                     /Tm_(binding_region)="57.4"
                     /Tm_(with_extension)="62.0"
                     /%GC_(binding_region)="50.0"
                     /%GC_(with_extension)="41.5"
                     /Hairpin_Tm_(with_extension)="46.8"
                     /Self_Dimer_Tm_(with_extension)="18.1"
                     /Pair_Dimer_Tm_(binding_region)="None"
                     /label="P2d PCR Product (78 R) - pJM220-mNeongreen not
                     codon optimized backbone piece"
     misc_feature    3113..>3124
                     /Color="#a6ccff"
                     /Transferred_From="BioBrick_suffix"
                     /Transferred_Similarity="100.00%"
                     /label="BioBrick suffix"
     ligation        3125..3143
                     /label="Ligation"
     misc_feature    3125..2102
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:2p-f9qmcqk"
                     /label="pJM220-T0T1::PrhaBAD-mS extraction PCR Product
                     (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     3125..3149
                     /Sequence="ATGAAATTCAACTAGTGCTCTGCAG"
                     /Tm="59.9"
                     /%GC="40.0"
                     /Hairpin_Tm="40.1"
                     /Self_Dimer_Tm="7.6"
                     /created_by="elinlarsson"
                     /note="Derived using Geneious Prime 2022.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pJM220-T0T1::PrhaBAD-mS extraction - 1.
                     pJM220-T0T1::pPhoA_stro extraction - 2c.
                     pJM220-T0T1::PrhaBAD-mN extraction - mNeonGreen - 3.
                     miniTn7(Gm)PA1/04/03-ecfp - lambda t0 terminator - 4.
                     miniTn7(Gm)PA1/04/03-ecfp - Pa1/04/03"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     pJM220_mS_F (<a
                     href=""urn:local:.:dw-eoqpkff"">pJM220-T0T1::PrhaBAD-mS
                     extraction - 1. pJM220-T0T1::pPhoA_stro extraction - 2c.
                     pJM220-T0T1::PrhaBAD-mN extraction - mNeonGreen - 3.
                     miniTn7(Gm)PA1/04/03-ecfp - lambda t0 terminator - 4.
                     miniTn7(Gm)PA1/04/03-ecfp - Pa1/04/03</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:fg-eor91yg"">pJM220_mS_F</a>"
                     /label="pJM220_mS_F"
     primer_bind     complement(3145..3164)
                     /Sequence="AGCTGTTCCTCCTTACTGCA"
                     /Tm="58.6"
                     /%GC="50.0"
                     /Hairpin_Tm="40.4"
                     /Self_Dimer_Tm="None"
                     /created_by="elinlarsson"
                     /label="term_P2d_RBS_R"
     misc_feature    3150..3157
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /label="RBS"
     misc_feature    3158..3164
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /label="spacer"
     gene            3165..3860
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /label="mScarlet-I"
     primer_bind     3165..3185
                     /Sequence="ATGAGTAAAGGAGAAGCTGTG"
                     /Tm="55.3"
                     /%GC="42.9"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /created_by="elinlarsson"
                     /label="mSc_F"
     primer_bind     complement(3836..3857)
                     /created_by="elinlarsson"
                     /Sequence="TTTGTATAGTTCATCCATGCCA"
                     /Tm="55.5"
                     /%GC="36.4"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /modified_by="elinlarsson"
                     /label="mSc_R"
     primer_bind     3861..3879
                     /Sequence="TCGCGAATCAGAACGCAGA"
                     /Tm="59.8"
                     /%GC="52.6"
                     /Hairpin_Tm="52.5"
                     /Self_Dimer_Tm="15.7"
                     /created_by="elinlarsson"
                     /label="phoA_bb_F"
     terminator      3866..4189
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /label="T1T2 terminator"
     misc_feature    4194..4196
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="MCS"
     primer          complement(4200..4219)
                     /note="sequence: TTCACTTATCTGGTTGGCCT"
                     /note="Derived using Geneious Prime 2022.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="eGRS37 -
                     pJM220-T0T1::PrhaBAD-mN-ASV_codonopt"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oGS43
                     (<a href=""urn:local:.:gv-el1szvh"">eGRS37 -
                     pJM220-T0T1::PrhaBAD-mN-ASV_codonopt</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oGS43 (<a
                     href=""urn:local:.:ar-eh1fbip"">pJM220-T0T1::PrhaBAD-mS</a
                     >)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oGS43 (<a href=""urn:local:.:or-el6a0pe"">5.
                     pJM220-T0T1::PrhaBAD-mS extraction (backbone piece eGRS38
                     or eGRS39)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oGS43 (<a href=""urn:local:.:n9-el68ttu"">eGRS36 -
                     backbone piece with Elowitz RBS - NZ_CP027755.1 -
                     Annotated P. synxantha 2-79 from pseudomonas.com
                     extraction (reversed) PCR Product</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oGS43 (<a
                     href=""urn:local:.:bb-eh1fjso"">pJM220-T0T1::PrhaBAD-mS</a
                     >)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oGS43 (<a href=""urn:local:.:hb-el1t76u"">eGRS37 -
                     backbone piece with Elowitz RBS and ASV-tag</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oGS43 (<a
                     href=""urn:local:.:an-eh1fbim"">pJM220-T0T1::PrhaBAD-mN</a
                     >)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oGS43 (<a href=""urn:local:.:d2-el1ow4c"">eGRS36 -
                     backbone piece with Elowitz RBS</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oGS43 (<a
                     href=""urn:local:.:aj-eh1fbii"">pJM220-T0T1::PrhaBAD-mN</a
                     >)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oGS43 (<a href=""urn:local:.:lk-el67ql8"">eGRS36 -
                     backbone piece with Elowitz RBS - NZ_CP027755.1 -
                     Annotated P. synxantha 2-79 from pseudomonas.com
                     extraction (reversed) PCR Product</a>)"
                     /label="oGS43"
     misc_feature    4205..4370
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="left end of Tn7"
     oriT            4522..4789
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /label="oriT"
     rep_origin      4810..5483
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /label="origin"
     gene            complement(5628..6488)
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /label="bla"
     CDS             complement(5631..6290)
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /label="AmpR"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
      361 tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga
      421 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
      481 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
      541 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ggttcgaggt cgacggtatc
      601 gataagctag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
      661 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      721 aatccaagct agactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
      781 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
      841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      901 agtaggacaa atccgccgct aggagcttgc ggcccggacg atcgaattgg ggatcttgaa
      961 gttcctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
     1021 ctgatgtgct taaaaactta ctcaatggaa ttctagatcg aattggccgc ggcgttgtga
     1081 caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt
     1141 ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata
     1201 gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag
     1261 cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg
     1321 ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct caaacctggg
     1381 cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat
     1441 gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta
     1501 ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac
     1561 ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt
     1621 tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc
     1681 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     1741 actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
     1801 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt
     1861 tacgaaccga acaggcttat gtcaattcga tccattgctg ttgacaaagg gaatcagggg
     1921 atcttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttcagagcgc
     1981 ttttgaagct aattcgatca tgcatgagct caaagagtgg aacaatgcag gacgccgtcc
     2041 ggtaacggtg ttgagcctcg gttgtgtggt gattattgtc gccgctaaca tcgtcggcat
     2101 cgagagcgct tttgaagcta attcgatcat gcatgagctc aaaagttaat cttttcaaca
     2161 gctgtcataa agttgtcacg gccgagactt atagtcgctt tgtttttatt ttttaatgta
     2221 ttaaagagga gaaattaagc atggtctcca agggcgagga agataatatg gcgtccctcc
     2281 ccgcgacgca tgaactccat atcttcgggt cgatcaacgg ggtggacttt gatatggtgg
     2341 ggcaagggac gggcaacccc aatgatggct acgaagagct caacctcaag agcacgaagg
     2401 gcgacctgca gtttagcccc tggatcctgg tgccgcacat cgggtatggc ttccatcaat
     2461 atctgcccta ccccgacggg atgtccccgt ttcaggcggc catggtcgac ggctcggggt
     2521 accaagtgca tcgcacgatg cagtttgagg atggcgcctc cctgaccgtg aactatcgct
     2581 acacgtacga aggcagccat atcaaggggg aagcgcaggt gaagggcacc ggctttccgg
     2641 cggacgggcc ggtgatgacc aattccctga cggccgcgga ttggtgtcgg tccaagaaga
     2701 cgtacccgaa cgataagacc atcatcagca cgttcaagtg gtcgtatacc accggcaacg
     2761 gcaagcgcta ccgttcgacg gcccgcacga cgtacacctt tgccaagccg atggcggcga
     2821 actatctgaa gaaccaaccc atgtatgtgt tccggaagac cgagctcaag cattcgaaga
     2881 cggagctgaa ttttaaggaa tggcagaagg cgtttaccga tgtgatgggg atggatgagc
     2941 tctataagta agactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt
     3001 gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatagac tagtgggtct
     3061 caggagttga cagctagctc agtcctaggg actatgctag ctactagaga cctactagta
     3121 gcggatgaaa ttcaactagt gctctgcagt aaggaggaac agctatgagt aaaggagaag
     3181 ctgtgattaa agagttcatg cgcttcaaag ttcacatgga gggttctatg aacggtcacg
     3241 agttcgagat cgaaggcgaa ggcgagggcc gtccgtatga aggcacccag accgccaaac
     3301 tgaaagtgac taaaggcggc ccgctgcctt tttcctggga catcctgagc ccgcaattta
     3361 tgtacggttc tagggcgttc atcaaacacc cagcggatat cccggactat tataagcagt
     3421 cttttccgga aggtttcaag tgggaacgcg taatgaattt tgaagatggt ggtgccgtga
     3481 ccgtcactca ggacacctcc ctggaggatg gcaccctgat ctataaagtt aaactgcgtg
     3541 gtactaattt tccacctgat ggcccggtga tgcagaaaaa gacgatgggt tgggaggcgt
     3601 ctaccgaacg cttgtatccg gaagatggtg tgctgaaagg cgacattaaa atggccctgc
     3661 gcctgaaaga tggcggccgc tatctggctg acttcaaaac cacgtacaaa gccaagaaac
     3721 ctgtgcagat gcctggcgcg tacaatgtgg accgcaaact ggacatcacc tctcataatg
     3781 aagattatac ggtggtagag caatatgagc gctccgaggg tcgtcattct accggtggca
     3841 tggatgaact atacaaataa tcgcgaatca gaacgcagaa gcggtctgat aaaacagaat
     3901 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     3961 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     4021 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     4081 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     4141 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcata aggccttgca
     4201 ggccaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt
     4261 agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta
     4321 aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt
     4381 tttcttcctg ttatgtttgg gcgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4441 gtttgcgtat tgggcgctct tccgcttcct cgctcactga cccgctgcgc tcggtcgttc
     4501 ggctgcggcg agcggtatca gagcttatcg gccagcctcg cagagcagga ttcccgttga
     4561 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
     4621 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
     4681 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
     4741 tgaccccgaa gcagggttat gcagcggaaa gtatacctta aggaatcccc tgataacgca
     4801 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4861 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcgagt
     4921 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4981 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5041 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5101 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5161 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5221 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5281 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5341 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5401 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5461 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5521 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5581 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5641 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5701 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5761 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5821 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5881 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5941 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6001 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6061 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6121 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6181 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6241 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6301 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6361 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6421 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6481 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6541 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6601 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6661 aataggcgta tcacgaggcc ctttcgtc
//
