LOCUS       sequence-201174-        5243 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses ShBle with FCP promoter/terminator, Golden Gate Assembly, 
            for Phaeodactylum transformation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5243)
  AUTHORS   Graff van Creveld S, Ben-Dor S, Mizrachi A, Alcolombri U, Hopes A, 
            Mock T, Rosenwasser S, Vardi A
  TITLE     Biochemical Characterization of a Novel Redox-Regulated Metacaspase 
            in a Marine Diatom.
  JOURNAL   Front Microbiol. 2021 Sep 8;12:688199. doi: 
            10.3389/fmicb.2021.688199. eCollection 2021.
  PUBMED    34566902
REFERENCE   2  (bases 1 to 5243)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Microbiol."; date: "2021-09-8"; volume: "12:688199. doi"; pages: " 
            10.3389/fmicb.2021.688199. eCollection 2021"
COMMENT     Sequence Label: pICH47732:PtFCP:ShBle
FEATURES             Location/Qualifiers
     source          1..5243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    76..100
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      192..823
                     /label=oriV
                     /note="incP origin of replication"
     promoter        910..1014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1015..1083,1084..1875)
                     /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     complement(1233..1252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2046..2634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2535..2554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2970..4118
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    4274..4298
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4589..4963)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
ORIGIN
        1 gtggggccca ctgcatccac cccagtacat taaaaacgtc cgcaatgtgt tattaagttg
       61 tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc
      121 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag tcagatcagg
      181 atctcctttg cgacgctcac cgggctggtt gccctcgccg ctgggctggc ggccgtctat
      241 ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg
      301 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
      361 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
      421 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
      481 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
      541 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
      601 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
      661 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
      721 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
      781 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
      841 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
      901 caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      961 ctcatgagac aataaccttg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1021 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1081 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1141 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1201 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1261 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1321 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1381 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1441 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1501 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1561 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1621 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1681 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt
     1741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2161 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2641 agcaacgcgg cctttttacg gttcctggca gatcctagat gtggcgcaac gatgccggcg
     2701 acaagcagga gcgcaccgac ttcttccgca tcaagtgttt tggctctcag gccgaggccc
     2761 acggcaagta tttgggcaag gggtcgctgg tattcgtgca gggcaagatt cggaatacca
     2821 agtacgagaa ggacggccag acggtctacg ggaccgactt cattgccgat aaggtggatt
     2881 atctggacac caaggcacca ggcgggtcaa atcaggaata agggcacatt gccccggcgt
     2941 gagtcggggc aatcccgcaa ggagggtgaa tgaatcggac gtttgaccgg aaggcataca
     3001 ggcaagaact gatcgacgcg gggttttccg ccgaggatgc cgaaaccatc gcaagccgca
     3061 ccgtcatgcg tgcgccccgc gaaaccttcc agtccgtcgg ctcgatggtc cagcaagcta
     3121 cggccaagat cgagcgcgac agcgtgcaac tggctccccc tgccctgccc gcgccatcgg
     3181 ccgccgtgga gcgttcgcgt cgtcttgaac aggaggcggc aggtttggcg aagtcgatga
     3241 ccatcgacac gcgaggaact atgacgacca agaagcgaaa aaccgccggc gaggacctgg
     3301 caaaacaggt cagcgaggcc aagcaggccg cgttgctgaa acacacgaag cagcagatca
     3361 aggaaatgca gctttccttg ttcgatattg cgccgtggcc ggacacgatg cgagcgatgc
     3421 caaacgacac ggcccgctct gccctgttca ccacgcgcaa caagaaaatc ccgcgcgagg
     3481 cgctgcaaaa caaggtcatt ttccacgtca acaaggacgt gaagatcacc tacaccggcg
     3541 tcgagctgcg ggccgacgat gacgaactgg tgtggcagca ggtgttggag tacgcgaagc
     3601 gcacccctat cggcgagccg atcaccttca cgttctacga gctttgccag gacctgggct
     3661 ggtcgatcaa tggccggtat tacacgaagg ccgaggaatg cctgtcgcgc ctacaggcga
     3721 cggcgatggg cttcacgtcc gaccgcgttg ggcacctgga atcggtgtcg ctgctgcacc
     3781 gcttccgcgt cctggaccgt ggcaagaaaa cgtcccgttg ccaggtcctg atcgacgagg
     3841 aaatcgtcgt gctgtttgct ggcgaccact acacgaaatt catatgggag aagtaccgca
     3901 agctgtcgcc gacggcccga cggatgttcg actatttcag ctcgcaccgg gagccgtacc
     3961 cgctcaagct ggaaaccttc cgcctcatgt gcggatcgga ttccacccgc gtgaagaagt
     4021 ggcgcgagca ggtcggcgaa gcctgcgaag agttgcgagg cagcggcctg gtggaacacg
     4081 cctgggtcaa tgatgacctg gtgcattgca aacgctaggg ccttgtgggg tcagttccgg
     4141 ctgggggttc agcagccagc gcctgatctg gggaaccctg tggttggcat gcacatacaa
     4201 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc
     4261 tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaaccact
     4321 tcgtgcagaa gacaattgca gcgctcgaga aaactcatcc tgtgccttcc tcactttctg
     4381 ctagtccact ccaacaatac ggcacgccgt gccctattcg gatattcttt ctctttccaa
     4441 aagttctagc tttggatgtt gaacttttat acaccatgaa taggcgagat tggcatccat
     4501 ggtgaagtcg agctagtgtt attcctgact gtgaaaccaa agcggagtga ctgcaactaa
     4561 tgaaaattaa atttttaagg aaggaagctc agtcctgctc ctcggccacg aagtgcacgc
     4621 agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg ccgatctcgg
     4681 tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac cactcggcgt
     4741 acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc accacctggt
     4801 cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg aagtcgtcct
     4861 ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt
     4921 cgcgcgcggt gagcaccgga acggcactgg tcaacttggc cattcttgac atctggcaac
     4981 cgtgaaatat gcgcgctgtg tatagtaagg aagccccagc tggagcgagc gtcgtagcgg
     5041 gaggccccca aaaggatacg tgttcaacca ggatttgctg tttacggcgt tcatgtcagg
     5101 atgtcgctct tgtgtcttat ccaggtccaa acagattgaa ttttgaatct ctccggatca
     5161 acgattactg tcagttgact gtgacagtgt agacgacgct gaaggtatgt ctccggatcc
     5221 gaattcggca ttgtcttcac aga
//
