LOCUS       sequence-408759-        6401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 Position 1 pUC19 vector with sgRNA and GmR. Used for 
            homologous recombination with Cas12a editing vector pC1.509. sgRNA 
            targets pC1.509 for removal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6401)
  AUTHORS   Victoria AJ, Selao TT, Moreno-Cabezuelo JA, Mills LA, Gale GAR, 
            Lea-Smith DJ, McCormick AJ
  TITLE     A toolbox to engineer the highly productive cyanobacterium 
            Synechococcus sp. PCC 11901.
  JOURNAL   Plant Physiol. 2024 May 7:kiae261. doi: 10.1093/plphys/kiae261.
  PUBMED    38713768
REFERENCE   2  (bases 1 to 6401)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol. 2024 May 7:kiae261. doi: 10.1093/plphys/kiae261."
COMMENT     Sequence Label: pC1.530
FEATURES             Location/Qualifiers
     source          1..6401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..171
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     236..255
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        538..572
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(823..1356)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1526..1531,1532..1548,1549..1554))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    2104..2128
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2284..3432)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3768..4356)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3848..3867)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4527..5318,5319..5387))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5150..5169
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5388..5492)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5579..6210
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6302..6326
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caatgccgaa ttcggatccg gagtgaccat ctcatctgta acatcattgg
       61 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
      121 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
      181 cagcatccat gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc
      241 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
      301 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
      361 taaatcgaac ttttgctgag ttgaaggatc agctcgagtt accaatgctt aatcagtgag
      421 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
      481 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataatgttg
      541 acagctagct cagtcctagg tataatgcta gcagctgatt taggcaaaaa cggaatttct
      601 actgttgtag atttaataaa tatagtttaa gtaaaacgaa tttctactgt tgtagatctc
      661 ggtaccaaat tccagaaaag aggcctcccg aaaggggggc cttttttcgt tttggtccag
      721 gtggtcgctt tcaggggtac cgagctcgaa ttggccgcgg cgttgtgaca atttaccgaa
      781 caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg
      841 gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc
      901 gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc
      961 atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac
     1021 tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca gaacgtaagc
     1081 cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact
     1141 acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc
     1201 tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc
     1261 gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact
     1321 gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     1381 cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aacaagccat
     1441 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     1501 tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg tcaagcttta
     1561 ctggagttat taacactgtg tacccggaca aacaccatta atcagcagga aggtaaattc
     1621 ttcaatatcc tgttcaacgg tcaactgttc ggtcagcagg cgataccaac aaaaaccaaa
     1681 aatcatatcc agcagcagtt cacgattgat atctttcggc agttcaccat tgctaatggc
     1741 atcttcaacc agttttttcg gtatctcacg acgacgttcc ataaactgat ctttcagttg
     1801 ggtcagggtt acagggtcca actgtgcttc tgcaataaca caacgaaatg cttcaccaca
     1861 aatggtttca cgccaaactt tccacagatt atgcagcaga aaatccagat cggctttaaa
     1921 gctacccaaa tccggaaatt tacgtacctg ttcgatttca ttttcataca cttcggcaat
     1981 cagtgctgct ttgttggtcc accaacgata aatggtcggt ttgcctgcac cggcgcgacg
     2041 tgccacgctt tcaatgctcg ctgcaattgt cttctgcacg aagtggttta aactatcagt
     2101 gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg
     2161 gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc
     2221 acagggttcc ccagatcagg cgctggctgc tgaaccccca gccggaactg accccacaag
     2281 gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct
     2341 gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga
     2401 atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga
     2461 gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat
     2521 gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg
     2581 gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga
     2641 ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag
     2701 gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag
     2761 ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac
     2821 ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt
     2881 cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt
     2941 gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc
     3001 cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg
     3061 tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct cgccggcggt
     3121 ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc
     3181 tgccgcctcc tgttcaagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc
     3241 agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga
     3301 gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc
     3361 ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa
     3421 cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg caatgtgccc
     3481 ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct tatcggcaat
     3541 gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc gaatcttgcc
     3601 ctgcacgaat accagcgacc ccttgcccaa atacttgccg tgggcctcgg cctgagagcc
     3661 aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg tcgccggcat cgttgcgcca
     3721 catctaggat ctgccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3781 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3841 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3901 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3961 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4021 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4081 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4141 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4201 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4261 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4321 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4381 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4441 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4501 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4561 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4621 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca
     4681 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4741 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4801 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4861 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4921 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4981 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5041 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5101 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5161 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5221 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5281 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5341 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5401 caatattatt gaagcattta tcaaggttat tgtctcatga gcggatacat atttgaatgt
     5461 atttagaaaa ataaacaaat aggggttccg cgcacgaatt ggccagcgct gccatttttg
     5521 gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag
     5581 cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac
     5641 agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag
     5701 acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct
     5761 gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca
     5821 agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc
     5881 agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg
     5941 ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc
     6001 tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat
     6061 ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc gcggtgtctc
     6121 gcacacggct tcgacggcgt ttctggcgcg tttgcagggc catagacggc cgccagccca
     6181 gcggcgaggg caaccagccc ggtgagcgtc gcaaaggaga tcctgatctg actgatgggc
     6241 tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg
     6301 gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc
     6361 ggacgttttt aatgtactgg ggtggatgca gtgggcccca c
//
