LOCUS       sequence-339549-        4204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L5 Integrase delivery plasmid to allow integration of L5 attP only 
            plasmids (e.g. plRL61, ID #163633)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4204)
  AUTHORS   Bosch B, DeJesus MA, Poulton NC, Zhang W, Engelhart CA, Zaveri A, 
            Lavalette S, Ruecker N, Trujillo C, Wallach JB, Li S, Ehrt S, Chait 
            BT, Schnappinger D, Rock JM
  TITLE     Genome-wide gene expression tuning reveals diverse vulnerabilities 
            of M. tuberculosis.
  JOURNAL   Cell. 2021 Jul 16. pii: S0092-8674(21)00824-2. doi: 
            10.1016/j.cell.2021.06.033.
  PUBMED    34297925
REFERENCE   2  (bases 1 to 4204)
  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: "Cell. 2021 
            Jul 16. pii: S0092-8674(21)00824-2. doi: 
            10.1016/j.cell.2021.06.033."
COMMENT     Sequence Label: plRL19
FEATURES             Location/Qualifiers
     source          1..4204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(436..1227,1228..1296))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1059..1078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1297..1401)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1469..1487
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1510..1515,1516..1533,1534..1540)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1546..1565
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     CDS             3262..3453
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3447..3469)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3555..3695
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3610..3629
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3710..3727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3881..4204,1..265))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3961..3980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
       61 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
      121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
      361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
      421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
      481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
      601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
      661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
      721 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
      781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
      841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
      961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1021 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     1081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     1441 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     1501 agcccaggct tgacacttta tgcttccggc tcgtataatg tgtggaattg tgagcgctca
     1561 caattcggat ccagcagagg agacctagtt ggcacgtcgc ggatggggat cgctgaagac
     1621 tcagcgcagc gggaggatcc aagcctcata cgtcaacccg caggacggtg tgaggtacta
     1681 cgcgctgcag acctacgaca acaagatgga cgccgaagcc tggctcgcgg gcgagaagcg
     1741 gctcatcgag atggagacct ggacccctcc acaggaccgg gcgaagaagg cagccgccag
     1801 cgccatcacg ctggaggagt acacccggaa gtggctcgtg gagcgcgacc tcgcagacgg
     1861 caccagggat ctgtacagcg ggcacgcgga gcgccgcatc tacccggtgc taggtgaagt
     1921 ggcggtcaca gagatgacgc cagctctggt gcgtgcgtgg tgggccggga tgggtaggaa
     1981 gcacccgact gcccgccggc atgcctacaa cgtcctccgg gcggtgatga acacagcggt
     2041 cgaggacaag ctgatcgcag agaacccgtg ccggatcgag cagaaggcag ccgatgagcg
     2101 cgacgtagag gcgctgacgc ctgaggagct ggacatcgtc gccgctgaga tcttcgagca
     2161 ctaccggatc gcggcataca tcctggcgtg gacgagcctc cggttcggag agctgatcga
     2221 gcttcgccgc aaggacatcg tggacgacgg catgacgatg aagctccggg tgcgccgtgg
     2281 cgcttcccgc gtggggaaca agatcgtcgt tggcaacgcc aagaccgtcc ggtcgaagcg
     2341 tcctgtgacg gttccgcctc acgtcgcgga gatgatccga gcgcacatga aggaccgtac
     2401 gaagatgaac aagggccccg aggcattcct ggtgaccacg acgcagggca accggctgtc
     2461 gaagtccgcg ttcaccaagt cgctgaagcg tggctacgcc aagatcggtc ggccggaact
     2521 ccgcatccac gacctccgcg ctgtcggcgc tacgttcgcc gctcaggcag gtgcgacgac
     2581 caaggagctg atggcccgtc tcggtcacac gactcctagg atggcgatga agtaccagat
     2641 ggcgtctgag gcccgcgacg aggctatcgc tgaggcgatg tccaagctgg ccaagacctc
     2701 ctgacgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc cagcagccgc
     2761 acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg
     2821 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     2881 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     2941 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     3001 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     3061 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     3121 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     3181 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     3241 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     3301 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     3361 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     3421 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3481 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3541 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     3601 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     3661 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3721 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3781 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3841 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3901 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3961 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4021 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4081 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4141 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4201 cggt
//
