LOCUS       sequence-422775-        3660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pUC19 containing the LL-FRT-erm-FRT-spacer sequence which serves as 
            template for BarSeq deletion constructs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3660)
  AUTHORS   Burgos HL, Burgos EF, Steinberger AJ, Suen G, Mandel MJ
  TITLE     Multiplexed Competition in a Synthetic Squid Light Organ Microbiome 
            Using Barcode-Tagged Gene Deletions.
  JOURNAL   mSystems. 2020 Dec 15;5(6):e00846-20. doi: 
            10.1128/mSystems.00846-20.
  PUBMED    33323415
REFERENCE   2  (bases 1 to 3660)
  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"; doi: "10.1128/mSystems"; 
            journalName: "mSystems"; date: "2020-12-15- 15"; volume: "5"; issue:
            "6"; pages: "e00846-20"
COMMENT     Sequence Label: pHB1
FEATURES             Location/Qualifiers
     source          1..3660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      303..891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     792..811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1045..1062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1179..1200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1215..1220,1221..1238,1239..1245)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1253..1269
                     /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     1258..1280
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1277..1293
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1277..1293
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1344..1377
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    2255..2288
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(2337..2354)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2337..2353)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2346..2368)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2562..2581)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2681..2703
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2741..2759)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2827..2931
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2932..3000,3001..3660,1..132)
                     /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(3150..3169)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
       61 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
      121 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
      181 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      241 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      301 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      361 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      421 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      481 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      541 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      601 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      661 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      721 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      781 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      841 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      901 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      961 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     1021 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     1081 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     1141 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     1201 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     1261 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tacgagacga
     1321 gcttcttata tatgcttcgc caggaagttc ctattctcta gaaagtatag gaacttcctt
     1381 agaagcaaac ttaagagtgt gttgatagtg cagtatctta aaattttgta taataggaat
     1441 tgaagttaaa ttagatgcta aaaatttgta attaagaagg agtgattaca tgaacaaaaa
     1501 tataaaatat tctcaaaact ttttaacgag tgaaaaagta ctcaaccaaa taataaaaca
     1561 attgaattta aaagaaaccg ataccgttta cgaaattgga acaggtaaag ggcatttaac
     1621 gacgaaactg gctaaaataa gtaaacaggt aacgtctatt gaattagaca gtcatctatt
     1681 caacttatcg tcagaaaaat taaaactgaa tactcgtgtc actttaattc accaagatat
     1741 tctacagttt caattcccta acaaacagag gtataaaatt gttgggagta ttccttacca
     1801 tttaagcaca caaattatta aaaaagtggt ttttgaaagc catgcgtctg acatctatct
     1861 gattgttgaa gaaggattct acaagcgtac cttggatatt caccgaacac tagggttgct
     1921 cttgcacact caagtctcga ttcagcaatt gcttaagctg ccagcggaat gctttcatcc
     1981 taaaccaaaa gtaaacagtg tcttaataaa acttacccgc cataccacag atgttccaga
     2041 taaatattgg aagctatata cgtactttgt ttcaaaatgg gtcaatcgag aatatcgtca
     2101 actgtttact aaaaatcagt ttcatcaagc aatgaaacac gccaaagtaa acaatttaag
     2161 taccgttact tatgagcaag tattgtctat ttttaatagt tatctattat ttaacgggag
     2221 gaaataatct agaatgcgag agtagggaac tgccgaagtt cctattctct agaaagtata
     2281 ggaacttcag cgctcatgca cttgattccg gatccccggg taccgagctc gaattcactg
     2341 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2401 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2461 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     2521 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     2581 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2641 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     2701 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     2761 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2821 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2881 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2941 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3001 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3061 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3121 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3181 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3241 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3301 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3361 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3421 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3481 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3541 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3601 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
//
