LOCUS       pETDuet-solo-His6NocO_D282A        6434 bp    DNA     circular UNA 25-MAY-2025
DEFINITION  Gibson Assembly ligation product.
ACCESSION   urn.local...8o-j7n26kb
VERSION     urn.local...8o-j7n26kb
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     source          join(<1412..6434,1..>100)
                     /organism="pETDuet-1"
                     /mol_type="other DNA"
                     /label="source pETDuet-1"
     misc_feature    2..29
                     /label="lacO"
     misc_feature    59..64
                     /label="T7_transl_en_RBS"
     ORF             71..1408
                     /Genetic_code="Standard"
                     /Reading_frame=3
                     /created_by="ORF Finder"
                     /modified_by="nateg"
                     /label="ORF 8 (frame 3)"
     misc_feature    79..1429
                     /Source="referenceToDocument: urn:local:.:95-j7n26kd"
                     /label="Not Nocuolin from Pedro PCR Product (concatenated
                     sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     ligation        79..100
                     /label="Ligation"
     misc_feature    complement(79..100)
                     /Mismatches=0
                     /%GC="54.5"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Pair_Dimer_Tm="None"
                     /Tm="61.6"
                     /annotation_group="67718930388280d8f91633-2d27-4e4e-b392-b
                     c17284c90b1"
                     /Sequence="GTGGTGATGATGGTGATGGCTG"
                     /created_by="primer3"
                     /Product_Size=5390
                     /label="pETDuet-MCS1-Gib-r"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    79..100
                     /label="His tag"
     misc_feature    79..106
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    101..106
                     /label="linker"
     CDS             <107..1405
                     /codon_start=1
                     /product="Uncharacterized protein"
                     /Original_Translation="MITSSNTSPISQINKVIPLLEKNYGNNLDSDYTEK
                     IQELDRNFNYSANSNVFWSEPEHSILYGTPLYEQASLSQKLALNHLHWVLFYKRVADS
                     ETELIHYNTITADCLMAANSDHKMLVELLAHETYQERQHIHAFSQINYKTTKALLGKK
                     ALVNPVSDETTVREQKKDISANYISTAANLLAMMLPGKKQSHADSDKYLSFPTNGFFN
                     GFSGKSDISRKQFFSDNWRSYPFLAANFYAVRYMANLLLKNHEFGIHCYFKKMHKNNE
                     FIASPTAISHYHFLDEAFHTTTSLFLGRDFHKYLPKPSRYEKWIANLAILQAQHENLN
                     AISGISTNRFLSDGDVIIFLMKLLQSPVFDMSAQEAIQWIEKCLCYEHEGFHVNLKVH
                     NKLLSELRKFAYQLDYLWPVNKEMRVMASGGSIKKAIKNNIKAFNQFSIAI"
                     /label="Uncharacterized protein CDS"
     source          <107..>1408
                     /organism="Nodularia sp. LEGE 06071 N071000019"
                     /label="source Nodularia sp. LEGE 06071 N071000019"
     gene            <107..1405
                     /product="Uncharacterized protein"
                     /modified_by="nateg"
                     /label="CylC"
     primer_bind     107..129
                     /Mismatches=0
                     /Extension="AGAGAGGGATCCG"
                     /%GC="34.8"
                     /Hairpin_Tm="46.4"
                     /Self_Dimer_Tm="15.5"
                     /Tm="53.4"
                     /annotation_group="522859378936845eb682d1a-8243-4538-85ab-
                     a2f7a5d0d3eb"
                     /Sequence="ATTACTAGCAGTAATACTTCACC"
                     /created_by="primer3"
                     /label="NocCylC-f"
     primer_bind     107..129
                     /Mismatches=0
                     /Extension="CAGCCATCACCATCATCACCACGGCAGC"
                     /%GC="34.8"
                     /Hairpin_Tm="46.4"
                     /Self_Dimer_Tm="15.5"
                     /Pair_Dimer_Tm="None"
                     /Tm="53.4"
                     /annotation_group="821550419631680d0ba9a7a-1946-4359-b307-
                     731a6a07bb8d"
                     /Sequence="ATTACTAGCAGTAATACTTCACC"
                     /created_by="primer3"
                     /Product_Size=1302
                     /label="MCS1-NodCylC-f"
     misc_feature    948..949
                     /Original_Bases="AT"
                     /label="AT"
                     /note="Geneious type: Editing History Replacement"
     Junction        959^960
                     /Reads_using=2
                     /Reads_supporting_discovery=1
                     /Junction_Type="Insertion"
                     /Insertion_Size=15
                     /Insertion="CTTCTCTATTTCTTG"
                     /ID="junction 1"
                     /label="CTTCTCTATTTCTTG"
     Junction        984^985
                     /Reads_using=2
                     /Reads_supporting_discovery=1
                     /Junction_Type="Insertion"
                     /Insertion_Size=25
                     /Insertion="CGACAACCACTTCTCTATTTCTTGG"
                     /ID="junction 1"
                     /label="CGACAACCACTTCTCTATTTCTTGG"
     misc_feature    complement(1380..1408)
                     /Mismatches=0
                     /Extension="ACACACCTGCAGTTA"
                     /%GC="24.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="3.2"
                     /Pair_Dimer_Tm="None"
                     /Tm="55.7"
                     /annotation_group="521998394602117b225014a-7cc8-4dd9-9598-
                     d3ffca055bb1"
                     /Sequence="TTAAATTGCTATAGAAAACTGATTAAAGG"
                     /created_by="primer3"
                     /Product_Size=1302
                     /label="NocCylC-r"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(1380..1408)
                     /Mismatches=0
                     /Extension="GTAGACGAGTCCATGTGCTTA"
                     /%GC="24.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="3.2"
                     /Pair_Dimer_Tm="None"
                     /Tm="55.7"
                     /annotation_group="821550419631680d0ba9a7a-1946-4359-b307-
                     731a6a07bb8d"
                     /Sequence="TTAAATTGCTATAGAAAACTGATTAAAGG"
                     /created_by="primer3"
                     /Product_Size=1302
                     /label="NodCylC-MCS2-r"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(1409..1411)
                     /label="stop"
     misc_feature    1409..1429
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    1412..100
                     /Source="referenceToDocument: urn:local:.:94-j7n26kd"
                     /label="pETDuet-1 PCR Product (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     ORF             <1412..1442
                     /codon_start=2
                     /created_by="nateg"
                     /label
     ligation        1412..1429
                     /label="Ligation"
     primer_bind     1412..1429
                     /Mismatches=0
                     /%GC="55.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="54.9"
                     /annotation_group="1469605865887063c36c131-60be-4338-8465-
                     2024fe90e782"
                     /Sequence="GCACATGGACTCGTCTAC"
                     /created_by="primer3"
                     /label="DuetCylA_down-f-2"
     misc_feature    complement(1412..1429)
                     /label="MCS2"
     misc_feature    complement(1462..1480)
                     /label="T7_Terminal_primer"
     gene            complement(2130..2990)
                     /gene="Ampicillin"
                     /label="Ampicillin"
     promoter        complement(3033..3061)
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="AmpR_promoter"
     rep_origin      3148..3767
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="pBR322_origin"
     misc_feature    4166..4188
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="pGEX_3_primer"
     misc_feature    complement(4182..4373)
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="ROP"
     misc_feature    complement(4945..6036)
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="lacI"
     gene            complement(6128..6379)
                     /gene="tet (887 - 636)"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
                     /label="tet (887 - 636)"
     misc_feature    6311..6330
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
                     /label="pBRrevBam_primer"
     promoter        6418..2
                     /created_by="nateg"
                     /label="T7 promoter"
ORIGIN      
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac ggcagcatta ctagcagtaa
      121 tacttcacca atatctcaaa ttaataaagt tatccctctc ttggaaaaga actatggaaa
      181 taaccttgac tcagactaca ctgaaaaaat ccaagaacta gatagaaact ttaactatag
      241 tgccaacagt aatgtttttt ggagtgaacc agaacactct atcctttatg gcacaccact
      301 ttatgaacaa gcttctctat ctcaaaaatt agcactcaat cacctgcatt gggtattgtt
      361 ttacaagaga gttgctgaca gtgaaactga gctaattcat tacaacacta tcacagcaga
      421 ttgtctcatg gctgcaaata gtgatcataa aatgctggtg gaactgttag ctcatgaaac
      481 ttaccaagaa cgtcaacata ttcatgcttt ttctcaaata aattataaaa ctactaaagc
      541 tttactaggt aaaaaagctc tagtgaatcc cgtttctgat gagactactg tacgggagca
      601 gaaaaaggat atatctgcca attatatatc cactgctgcc aacttgctcg caatgatgct
      661 cccaggtaaa aagcaatctc atgccgattc tgataaatat ttgtcatttc ctaccaacgg
      721 attttttaat ggattttcag gaaaatcaga tatatcaaga aaacagttct tttcagataa
      781 ttggagaagt tatccattct tggctgctaa tttttatgct gtgcgctaca tggctaatct
      841 actattgaaa aaccatgaat ttggtataca ttgctacttc aaaaaaatgc ataaaaataa
      901 tgaatttata gcgagtccaa cagctatatc tcactatcat tttttagcgg aagcttttca
      961 tacaaccact tctctatttc ttggtcgaga ctttcataaa tatttaccaa agccatcaag
     1021 atatgaaaaa tggattgcca acttagctat cttacaggca cagcacgaaa acctcaatgc
     1081 tatttctgga atatctacca accgcttttt gagtgatggt gatgtgataa tttttctgat
     1141 gaaactcttg cagagtcctg tatttgatat gtcggctcag gaagcaattc agtggattga
     1201 aaagtgccta tgttatgagc atgaaggatt tcatgtcaac ctgaaggtac ataataaact
     1261 gttatcagaa ttacgtaagt ttgcatatca actagactac ttgtggcctg taaataagga
     1321 aatgcgtgta atggcttcag gaggttcaat taagaaagcg atcaaaaata atatcaaagc
     1381 ctttaatcag ttttctatag caatttaata agcacatgga ctcgtctact agcgcagctt
     1441 aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1501 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga
     1561 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1621 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1681 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1741 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1801 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1861 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1921 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1981 cgcgaatttt aacaaaatat taacgtttac aatttctggc ggcacgatgg catgagatta
     2041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2161 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2221 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2281 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2341 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2401 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2461 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2521 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2581 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2641 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2701 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2761 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2821 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2881 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2941 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3001 tttcaatcat gattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3061 atgtatttag aaaaataaac aaataggtca tgaccaaaat cccttaacgt gagttttcgt
     3121 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3181 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3241 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3301 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3361 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3421 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3481 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3541 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3601 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3661 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3721 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3781 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3841 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3901 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3961 cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga
     4021 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     4081 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4141 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4201 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     4261 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     4321 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     4381 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     4441 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     4501 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     4561 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     4621 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     4681 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     4741 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     4801 cgggtcctca acgacaggag cacgatcatg ctagtcatgc cccgcgccca ccggaaggag
     4861 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     4921 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4981 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     5041 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     5101 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     5161 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     5221 cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     5281 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     5341 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     5401 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     5461 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     5521 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     5581 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     5641 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     5701 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     5761 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     5821 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     5881 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     5941 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     6001 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     6061 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     6121 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     6181 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     6241 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     6301 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     6361 tgccggccac gatgcgtccg gcgtagagga tcgagatcga tctcgatccc gcgaaattaa
     6421 tacgactcac tata
//
