LOCUS       pCDFDuet-His6NocM-emptyMCS2        4042 bp    DNA     circular UNA 30-JUN-2023
DEFINITION  Gibson Assembly ligation product.
ACCESSION   urn.local...783-gkvujo9
VERSION     urn.local...783-gkvujo9
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     ORF             4013..288
                     /Genetic_code="Standard"
                     /Reading_frame=1
                     /created_by="ORF Finder"
                     /modified_by="nateg"
                     /label="ORF 7 (frame 1)"
     misc_feature    4021..310
                     /Source="referenceToDocument: urn:local:.:78e-gkvujoc"
                     /label="insert CylB (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    4021..6
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    4021..4042
                     /label="His6"
     misc_feature    1..6
                     /label="linker"
     gene            <7..288
                     /created_by="nateg"
                     /label="CylB"
     source          <7..>288
                     /organism="Nodularia sp. LEGE 06071 N071000019"
                     /label="source Nodularia sp. LEGE 06071 N071000019"
     primer_bind     7..27
                     /Mismatches=0
                     /Extension="CAGCCATCACCATCATCACCACGGCAGC"
                     /%GC="38.1"
                     /Hairpin_Tm="35.6"
                     /Self_Dimer_Tm="None"
                     /Pair_Dimer_Tm="None"
                     /Tm="56.1"
                     /annotation_group="146605817781496d8974faa-06c5-4283-be8f-
                     7282aca2b758"
                     /Sequence="ACGCAACTTTTTGAAGACAAC"
                     /created_by="primer3"
                     /Product_Size=282
                     /label="pETDuet1-NodCylB-f"
     misc_feature    complement(265..288)
                     /Mismatches=0
                     /Extension="CAAGCTTGTCGACCTGCAGTTA"
                     /%GC="37.5"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Pair_Dimer_Tm="None"
                     /Tm="57.0"
                     /annotation_group="146605817781496d8974faa-06c5-4283-be8f-
                     7282aca2b758"
                     /Sequence="TTAGTTTTCCTCAGCCACATATTC"
                     /created_by="primer3"
                     /Product_Size=282
                     /label="NodCylB-DuetMCS1-r"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    289..310
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    complement(289..291)
                     /label="Stop"
     misc_feature    292..4042
                     /Source="referenceToDocument: urn:local:.:78d-gkvujoc"
                     /label="pCDFDuet-1-MCS1-Gib (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     ligation        292..310
                     /label="Ligation"
     misc_feature    complement(292..310)
                     /label="MCS1"
     primer_bind     292..310
                     /Mismatches=0
                     /%GC="57.9"
                     /Hairpin_Tm="31.9"
                     /Self_Dimer_Tm="24.8"
                     /Pair_Dimer_Tm="None"
                     /Tm="59.1"
                     /annotation_group="820846052440565c278bb59-9110-4bba-8a5e-
                     2e2643b6b6ad"
                     /Sequence="CTGCAGGTCGACAAGCTTG"
                     /created_by="primer3"
                     /Product_Size=3751
                     /label="Duet-MCS1-Gib-f"
     promoter        375..391
                     /vntifkey=30
                     /label="T7\promoter"
     protein_bind    396..416
                     /vntifkey=31
                     /label="lacO"
     misc_feature    complement(419..458)
                     /%GC="27.5"
                     /Sequence="GTATATCTCCTTCTTATACTTAACTAATATACTAAGATGG"
                     /Mismatches=0
                     /Extension="G"
                     /annotation_group="eb3c08f3-1a0b-4291-95cf-25c1e6d720c4"
                     /Product_Size=3681
                     /label="DuetCylA_up-r"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(419..458)
                     /%GC="27.5"
                     /Sequence="GTATATCTCCTTCTTATACTTAACTAATATACTAAGATGG"
                     /Mismatches=0
                     /Extension="G"
                     /annotation_group="9035a987-f8a9-42ca-a8ac-af7708b06283"
                     /Product_Size=3942
                     /label="DuetCylA_up-r"
                     /note="Geneious type: primer_bind_reverse"
     RBS             447..452
                     /vntifkey=32
                     /label="RBS"
     CDS             461..589
                     /vntifkey=4
                     /label="MCS-2\ORF"
     protein_bind    527..571
                     /vntifkey=31
                     /label="S-Tag"
     primer_bind     559..576
                     /%GC="55.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="54.9"
                     /created_by="primer3"
                     /Sequence="GCACATGGACTCGTCTAC"
                     /Mismatches=0
                     /annotation_group="eb3c08f3-1a0b-4291-95cf-25c1e6d720c4"
                     /Product_Size=3681
                     /label="DuetCylA_down-f-2"
     primer_bind     559..576
                     /%GC="55.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="54.9"
                     /created_by="primer3"
                     /Sequence="GCACATGGACTCGTCTAC"
                     /Mismatches=0
                     /annotation_group="9035a987-f8a9-42ca-a8ac-af7708b06283"
                     /Product_Size=3942
                     /label="DuetCylA_down-f-2"
     primer          complement(609..627)
                     /vntifkey=27
                     /label="T7\Terminator\primer"
     terminator      623..670
                     /vntifkey=43
                     /label="T7\Terminator"
     CDS             complement(844..1632)
                     /vntifkey=4
                     /label="aadA\(Correct,\original\source\Tn21)"
     RBS             complement(1638..1642)
                     /vntifkey=32
                     /label="RBS"
     -10_signal      complement(1674..1680)
                     /vntifkey=47
                     /label="-10\Ap"
     -35_signal      complement(1697..1702)
                     /vntifkey=48
                     /label="-35\Ap"
     rep_origin      complement(1772..2510)
                     /vntifkey=33
                     /label="CloDF13\ori"
     rep_origin      1863
                     /vntifkey=33
                     /label="Replication\Origin"
     promoter        2315..2344
                     /vntifkey=30
                     /label="RNA\I\promoter"
     promoter        2315..2320
                     /vntifkey=30
                     /note="-35 region (RNA I)"
                     /label="Promoter_P_5"
     promoter        2339..2344
                     /vntifkey=30
                     /note="-10 region (RNA I)"
                     /label="Promoter_P_6"
     terminator      complement(2427..2448)
                     /vntifkey=43
                     /note="put. stem-loop structure; put. transcription
                     terminator (RNA I)"
                     /label="Terminator_1"
     repeat_unit     complement(2427..2435)
                     /vntifkey=35
                     /note="inverted repeat A'"
                     /label="Repeat_Unit_1"
     repeat_unit     complement(2440..2448)
                     /vntifkey=35
                     /note="inverted repeat A"
                     /label="Repeat_Unit_2"
     promoter        complement(2467..2495)
                     /vntifkey=30
                     /label="RNA\II\Promoter"
     promoter        complement(2467..2472)
                     /vntifkey=30
                     /note="-10 region (RNA II)"
                     /label="Promoter_P_7"
     terminator      complement(2484..2506)
                     /vntifkey=43
                     /note="put. stem-loop structure; put. transcription
                     terminator (cex gene)"
                     /label="Terminator_2"
     repeat_unit     complement(2484..2491)
                     /vntifkey=35
                     /note="inverted repeat B'"
                     /label="Repeat_Unit_3"
     promoter        complement(2490..2495)
                     /vntifkey=30
                     /note="-35 region (RNA II)"
                     /label="Promoter_P_8"
     repeat_unit     complement(2499..2506)
                     /vntifkey=35
                     /note="inverted repeat B"
                     /label="Repeat_Unit_4"
     CDS             complement(2720..3802)
                     /vntifkey=4
                     /note="Wildtype lacI gene"
                     /label="lacI"
     promoter        3926..3942
                     /vntifkey=30
                     /label="T7\Promoter"
     protein_bind    3947..3967
                     /vntifkey=31
                     /label="lacO"
     RBS             4000..4005
                     /vntifkey=32
                     /label="RBS-1"
     ligation        4021..4042
                     /label="Ligation"
     misc_feature    complement(4021..4042)
                     /Mismatches=0
                     /%GC="54.5"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Pair_Dimer_Tm="None"
                     /Tm="61.6"
                     /annotation_group="820846052440565c278bb59-9110-4bba-8a5e-
                     2e2643b6b6ad"
                     /Sequence="GTGGTGATGATGGTGATGGCTG"
                     /created_by="primer3"
                     /Product_Size=3751
                     /label="Duet-MCS1-Gib-r"
                     /note="Geneious type: primer_bind_reverse"
     misc_binding    4025..4042
                     /vntifkey=20
                     /label="His-Tag"
ORIGIN      
        1 ggcagcacgc aactttttga agacaacaaa gcaagtgaac agagcgagca atttacacaa
       61 ctaaaaatta gcgagtggat tgttgactat atgtcagatt tactagaaat tgaacccgat
      121 gaagtagatg tagaaactac tttcgcgcgt tatggtttag attcatcagc cgcagtgatt
      181 ttaacgggag atttaggaaa ttggttagga aaagagatag aaccaacagt catgtatgat
      241 tacccaacaa tctccaagtt agcagaatat gtggctgagg aaaactaata actgcaggtc
      301 gacaagcttg cggccgcata atgcttaagt cgaacagaaa gtaatcgtat tgtacacggc
      361 cgcataatcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
      421 atcttagtat attagttaag tataagaagg agatatacat atggcagatc tcaattggat
      481 atcggccggc cacgcgatcg ctgacgtcgg taccctcgag tctggtaaag aaaccgctgc
      541 tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc
      601 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
      661 gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt
      721 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
      781 gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca
      841 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
      901 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
      961 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
     1021 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
     1081 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
     1141 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
     1201 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
     1261 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
     1321 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
     1381 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
     1441 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
     1501 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
     1561 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac
     1621 cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1681 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagcta gctcactcgg
     1741 tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa agttacccac
     1801 agattccgtg gataagcagg ggactaacat gtgaggcaaa acagcagggc cgcgccggtg
     1861 gcgtttttcc ataggctccg ccctcctgcc agagttcaca taaacagacg cttttccggt
     1921 gcatctgtgg gagccgtgag gctcaaccat gaatctgaca gtacgggcga aacccgacag
     1981 gacttaaaga tccccaccgt ttccggcggg tcgctccctc ttgcgctctc ctgttccgac
     2041 cctgccgttt accggatacc tgttccgcct ttctccctta cgggaagtgt ggcgctttct
     2101 catagctcac acactggtat ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta
     2161 agcaagaact ccccgttcag cccgactgct gcgccttatc cggtaactgt tcacttgagt
     2221 ccaacccgga aaagcacggt aaaacgccac tggcagcagc cattggtaac tgggagttcg
     2281 cagaggattt gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa agtccggcta
     2341 cactggaagg acagatttgg ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag
     2401 ttccccaact gacttaacct tcgatcaaac cacctcccca ggtggttttt tcgtttacag
     2461 ggcaaaagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctactg
     2521 aaccgctcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
     2581 tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac
     2641 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     2701 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     2821 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     2881 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     2941 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     3001 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     3061 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     3121 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     3181 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     3241 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     3301 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     3361 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     3421 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     3481 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     3541 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     3601 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     3661 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     3721 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     3781 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     3841 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     3901 ttatgcgact cctgcattag gaaattaata cgactcacta taggggaatt gtgagcggat
     3961 aacaattccc ctgtagaaat aattttgttt aactttaata aggagatata ccatgggcag
     4021 cagccatcac catcatcacc ac
//
