LOCUS       sequence-169710-        7030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ShBle with FCP promoter/terminator for F. cylindrus 
            transformation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7030)
  AUTHORS   Mock lab
  TITLE     Mock lab F. cylindrus plasmids
REFERENCE   2  (bases 1 to 7030)
  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"
COMMENT     Sequence Label: pICH47732:FCP:ShBle
FEATURES             Location/Qualifiers
     source          1..7030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     71..93
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     85..102
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     86..102
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1108..1482
                     /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"
     primer_bind     complement(2628..2644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2628..2644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2641..2663)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2652..2668
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    2733..2757
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2913..4061)
                     /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(4397..4985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4477..4496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5156..5947,5948..6016))
                     /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     5779..5798
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6017..6121)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6208..6839
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6931..6955
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caatgccgaa ttcggatccg gaggctgcaa ggcgattaag ttgggtaacg
       61 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcga gctcggtacc
      121 ccaaagtaag gcatagaaat aatctgcatg aaaagatcta acgatttcta tcagttcccc
      181 tacaaatgga tgcttagggg tgcttgtatg agtgtacgaa agagccaacc aaaaaataat
      241 tttttattct gatttttaat tttttgtgct aaaacatgct gcgctgtgta cttcctccgt
      301 gcaccggtaa acgcgcacgg aggaaataca gtgttttaga aaaacaacgc gcaaaaatgc
      361 caatctttat tattttgaag tgaagcgcgc tatttactgt tatcgcgcta atactgtaaa
      421 aattcgcgct gatcttgatg ataccacctt aactttgttt ctattttttg ataacaaata
      481 caaaatctac tagtattgtg caccctccta ctactagcta ctactaggga gtgtcgtact
      541 agtagtctaa aaatacgcgt gttcaacatg cgcgtgctag aagtatgatg cgtgtgtaac
      601 aaatttttaa aagattaaaa aaatctttat tttgagggat tggatgcgtg atagaatgga
      661 gaattgtgtt cgtgggattc attttgacct accaattaaa acgaataaaa atgtcagcgt
      721 cgattataga cacgaagtac agaccagaca gcgcgtcgac gtcagatttt tatggccaat
      781 cagattgttt ccaacacaca aatggaaaga aagaatgcat tctgtggaag agtgtacatc
      841 cgggggaatt tagaacgaaa accacgtcgg tgaattgtac acgcgacaat cctccaatcg
      901 tggataatcc aggatttttt gttgatgttg aactgcatat ttgccgatcc ttcatgcgaa
      961 ctccaaattt ctatatcatt ttttcagcaa aatccaatat ttttcagcaa aatccaaaaa
     1021 acaagccatc gttttaccgc tttcgatctt ctcataactt tattctttca acagtataaa
     1081 ctaccaaaaa caaacttata tatcaaaatg gccaagttga ccagtgccgt tccggtgctc
     1141 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     1201 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     1261 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     1321 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     1381 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     1441 gccggcaact gcgtgcactt cgtggccgag gagcaggact gagcatttta ttaatcctta
     1501 tttgatcgat tcattgattc gaatatagtt agacccttcc gttgacaact acaatcgatt
     1561 cctaatttaa ttatttagta gagttccttg agtttttgtc acaaatatta ttatctactt
     1621 ctatcatata ctactattca tagctagcta cgcattgttt cctcaatcca tacgcagttg
     1681 tactgttaca ttcacaaccc gtatccctca tgacctcttg actgtatcct acagtacgta
     1741 tttcctttat ttttctcttc cacgagctac agtagccaca cccctacggt acggtacagt
     1801 attgataaat caaccgacga cagcgcctct caactgccaa acagctggaa tgcgttcgtt
     1861 tgagaatgac cccaagacaa aagatccatt ttcaacctac agtaacgtcc gcccacgttt
     1921 gcaggatcca aaaatgcgag gaatcaatcc agtcgaacta gtcgagaaag ggcaacaaag
     1981 atccaatttc cgtactagac agatgtacag tatggaatcg agtgaatcaa accagtcgat
     2041 tcagtcgaac tagtcgagga ttgtaatcaa tactgtacat agtttgcgct ttgccttcca
     2101 ctaagttacc ggtacaggag tagtagattt tgtgagattt ttcatgacaa acaaatcgaa
     2161 aagtaaaaaa aaaacctatt agaaatcaac aatgacgacg atgatgatgt aacatgatga
     2221 ggaggataat cataatagtg acaattcaac tatcgtatca ccttatttct tcacaatcag
     2281 ccaaaagaaa attccctaga ggtcagcaga aaaaattacg aaaagaacgg cgtactgtac
     2341 agcaacggcg aaggattgac aaatgaatag tattattaga ataaaaagtg tttggttttc
     2401 gttatcgtta tatttatcgg tattgttact attactattt agtttggttc tattttttaa
     2461 tagtaataat aataataata atgatacaac aatcatagta gtgggcgtta atggatttct
     2521 tctaccaaca agaacaacaa taacgccgac atcagcacag cgactactgt tgtcgtctac
     2581 tacagcacaa caacaacgac taggcatgca agcttggcgt aatcatggtc atagctgttt
     2641 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgc tgcaattgtc
     2701 ttctgcacga agtggtttaa actatcagtg tttgacagga tatattggcg ggtaaaccta
     2761 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc
     2821 gttcgtccat ttgtatgtgc atgccaacca cagggttccc cagatcaggc gctggctgct
     2881 gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac caggtcatca
     2941 ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc ttcgccgacc
     3001 tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc
     3061 agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc
     3121 ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc
     3181 acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc acggtccagg
     3241 acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag
     3301 cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta ataccggcca
     3361 ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg
     3421 ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc
     3481 cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg
     3541 ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc
     3601 gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa caaggaaagc
     3661 tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg
     3721 acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat agttcctcgc
     3781 gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcaagacg acgcgaacgc
     3841 tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct gtcgcgctcg
     3901 atcttggccg tagcttgctg gaccatcgag ccgacggact ggaaggtttc gcggggcgca
     3961 cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc
     4021 agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt
     4081 gccccgactc acgccggggc aatgtgccct tattcctgat ttgacccgcc tggtgccttg
     4141 gtgtccagat aatccacctt atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc
     4201 ttctcgtact tggtattccg aatcttgccc tgcacgaata ccagcgaccc cttgcccaaa
     4261 tacttgccgt gggcctcggc ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc
     4321 tcctgcttgt cgccggcatc gttgcgccac atctaggatc tgccaggaac cgtaaaaagg
     4381 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4441 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4501 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4561 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4621 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4681 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4741 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4801 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4861 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4921 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4981 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5041 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5101 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5161 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5221 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5281 ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc
     5341 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5401 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5461 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5521 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5581 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5641 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5701 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5761 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5821 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5881 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5941 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6001 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat caaggttatt
     6061 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6121 gcacgaattg gccagcgctg ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag
     6181 cccctggggg gatgggaggc ccgcgttagc gggccgggag ggttcgagaa gggggggcac
     6241 cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta
     6301 taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga
     6361 aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg
     6421 cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag
     6481 tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca
     6541 tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc
     6601 acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg
     6661 cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt
     6721 atccacaacg ccggcggccg cggtgtctcg cacacggctt cgacggcgtt tctggcgcgt
     6781 ttgcagggcc atagacggcc gccagcccag cggcgagggc aaccagcccg gtgagcgtcg
     6841 caaaggagat cctgatctga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
     6901 ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaatt
     6961 gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg gtggatgcag
     7021 tgggccccac
//
