LOCUS       sequence-315537-        5371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of PilB L37 Amber with a C-terminal histag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5371)
  AUTHORS   Katrina Forest
  TITLE     Forest Lab PilB Plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5371)
  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: 
            "unpublished"
COMMENT     Sequence Label: pUCP20 PilBL37*
FEATURES             Location/Qualifiers
     source          1..5371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             3925..3942
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3942..4775)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      4789..5140
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     primer_bind     complement(5202..5221)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5321..5343
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 aacgacagca tccaactgag cggtctgtcc cgacagctcg tccaagccaa tctactcgac
     2281 gaaaagaccg ccctgcaggc ccagacgcag gcgcagcgca acaagtagtc gctggtgacg
     2341 cacctggtgc agaacaagct ggtaagcggc ctggccctgg ccgaattgtc cgccgagcag
     2401 ttcggcatcg cctattgcga cctgaacagc ctggacaggg aaagctttcc gcgggacgcc
     2461 atcagcgaaa agctggtccg ccagcaccgg gtcattcccc tgtggcggcg gggcaacaag
     2521 ttgttcgtag gcatctccga cgccgccaac caccaggcca tcaacgacgt ccagttcagc
     2581 actggcctga cgaccgaggc cattctggtc gaggacgaca agctcggcct tgccatcgac
     2641 aagctattcg agaatgcaac cgacggcttg gcaggtctcg atgacgtgga cctggaagga
     2701 ctggatgtag gcgtcaagga aacgtccgga caagaagata ctggcgcgga ggcagacgac
     2761 gctcccgtcg tccgcttcgt caacaagatg ctgctggacg ctatcaaagg cgggtcttcc
     2821 gacttgcact tcgagcccta cgagaagatc taccgggtgc gcttccgtac cgacggcatg
     2881 cttcacgaag tggccaagcc gccgatccag ttggccagtc gtatctctgc tcgtctcaag
     2941 gtaatggccg gcctggatat ctccgaacga cgcaagccgc aggacggacg gatcaagatg
     3001 cgcgtgtcga agaccaagtc catcgacttc cgcgtcaaca ccctgccgac cctgtggggc
     3061 gagaagatcg tgatgcggat cctcgactcc tccagcgcgc agatgggcat cgacgccctg
     3121 ggctacgagg aggaccagaa ggaactctac ctggccgcac tcaagcagcc gcagggcatg
     3181 atcctggtca ccggccccac cggctcgggc aagacggtat cgctgtacac cggcctgaac
     3241 atcctcaata ccaccgacat caacatttcc accgccgaag acccggtgga gatcaacctg
     3301 gaaggcatca accaggtcaa cgtcaatccg cgccagggca tggacttctc ccaggcgctg
     3361 cgcgccttcc tgcgccagga cccggacgtg atcatggtcg gcgagatccg cgacctggag
     3421 accgccgaga tcgccatcaa ggcggcgcag accgggcata tggtgatgtc caccctgcac
     3481 accaacagcg ccgccgagac cctgacccgc ctgctgaaca tgggcgtacc ggcgttcaac
     3541 ctggcgacct cggtgaacct gatcatcgcc cagcgccttg cgcgaaaact ctgttcgcac
     3601 tgcaagaaag agcacgacgt accgaaggaa accctgcttc acgaaggttt cccagaagag
     3661 ttgatcggca ccttcaagct gtattcacca gtgggctgcg accattgcaa gaacggttac
     3721 aagggccgtg tcggtattta tgaagtagtt aaaaacaccc cggccctgca gcggattatc
     3781 atggaggaag gcaactccat cgagatcgcc gagcaagccc gcaaagaagg cttcaacgat
     3841 ctgcgcacct caggcctgct gaaagccatg caggggatca ccagcctgga ggaagtcaac
     3901 cgcgtgacca aggataggcc gggccatcat catcaccatc attaagccag atccagcggc
     3961 atctgggtta gtcgagcgcg ggccgcttcc catgtctcac cagggcgagc ctgtttcgcg
     4021 atctcagcat ctgaaatctt cccggccttg cgcttcgctg gggccttacc caccgccttg
     4081 gcgggcttct tcggtccaaa actgaacaac agatgtgtga ccttgcgccc ggtctttcgc
     4141 tgcgcccact ccacctgtag cgggctgtgc tcgttgatct gcgtcacggc tggatcaagc
     4201 actcgcaact tgaagtcctt gatcgaggga taccggcctt ccagttgaaa ccactttcgc
     4261 agctggtcaa tttctatttc gcgctggccg atgctgtccc attgcatgag cagctcgtaa
     4321 agcctgatcg cgtgggtgct gtccatcttg gccacgtcag ccaaggcgta tttggtgaac
     4381 tgtttggtga gttccgtcag gtacggcagc atgtctttgg tgaacctgag ttctacacgg
     4441 ccctcaccct cccggtagat gattgtttgc acccagccgg taatcatcac actcggtctt
     4501 ttccccttgc cattgggctc ttgggttaac cggacttccc gccgtttcag gcgcagggcc
     4561 gcttctttga gctggttgta ggaagattcg atagggacac ccgccatcgt cgctatgtcc
     4621 tccgccgtca ctgaatacat cacttcatcg gtgacaggct cgctcctctt cacctggcta
     4681 atacaggcca gaacgatccg ctgttcctga acactgaggc gatacgcggc ctcgaccagg
     4741 gcattgcttt tgtaaaccat tgggggtgag gccacgttcg acattccttg tgtataaggg
     4801 gacactgtat ctgcgtccca caatacaaca aatccgtccc tttacaacaa caaatccgtc
     4861 ccttcttaac aacaaatccg tcccttaatg gcaacaaatc cgtccctttt taaactctac
     4921 aggccacgga ttacgtggcc tgtagacgtc ctaaaaggtt taaaagggaa aaggaagaaa
     4981 agggtggaaa cgcaaaaaac gcaccactac gtggccccgt tggggccgca tttgtgcccc
     5041 tgaaggggcg ggggaggcgt ctgggcaatc cccgttttac cagtccccta tcgccgcctg
     5101 agagggcgca ggaagcgagt aatcagggta tcgaggcgga ttcacccttg gcgtccaacc
     5161 agcggcacca gcggcgcctg agaggtatgg tgcactctca gtacaatctg ctctgatgcc
     5221 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5281 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5341 aggttttcac cgtcatcacc gaaacgcgcg a
//
