LOCUS       sequence-408096-        4727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing integrase for LPattB7 site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4727)
  AUTHORS   Triassi AJ, Fields BD, Monahan CE, Means JM, Park Y, Doosthosseini 
            H, Padmakumar JP, Isabella VM, Voigt CA
  TITLE     Redesign of an Escherichia coli Nissle treatment for phenylketonuria
            using insulated genomic landing pads and genetic circuits to reduce 
            burden.
  JOURNAL   Cell Syst. 2023 Jun 21;14(6):512-524.e12. doi: 
            10.1016/j.cels.2023.05.004.
  PUBMED    37348465
REFERENCE   2  (bases 1 to 4727)
  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: 
            "10.1016/j.cels.2023.05"; journalName: "Cell Syst"; date: 
            "2023-06-21- 21"; volume: "14"; issue: "6"; pages: "512-524.e12"
COMMENT     Sequence Label: plYJP022-int7
FEATURES             Location/Qualifiers
     source          1..4727
                     /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      1470..1826
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            1877..1985
                     /note="incP origin of transfer"
     primer_bind     2226..2243
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2360..2381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2396..2401,2402..2419,2420..2426)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2434..2450
                     /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     2439..2461
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2458..2474
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2458..2474
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2519..2553
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     misc_RNA        2561..2610
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     primer_bind     complement(4333..4350)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4333..4349)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4342..4364)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4558..4577)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4677..4699
                     /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 aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagccggtt gtcagccgtt aagtgttcct gtgtcactca
     1501 aaattgcttt gagaggctct aagggcttct cagtgcgtta catccctggc ttgttgtcca
     1561 caaccgttaa accttaaaag ctttaaaagc cttatatatt cttttttttc ttataaaact
     1621 taaaacctta gaggctattt aagttgctga tttatattaa ttttattgtt caaacatgag
     1681 agcttagtac gtgaaacatg agagcttagt acgttagcca tgagagctta gtacgttagc
     1741 catgagggtt tagttcgtta aacatgagag cttagtacgt taaacttgag agcttagtac
     1801 gtgaaacatg agagcttagt acgtactatc aacaggttga actgcccatg ttctttcctg
     1861 cgttatcaga gcttatcggc cagcctcgca gagcaggatt cccgttgagc accgccaggt
     1921 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
     1981 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
     2041 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
     2101 agggttatgc agcggaaagt ataccttaaa tgttctttcc tgcgttatcc cctgattctg
     2161 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     2221 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     2281 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     2341 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     2401 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     2461 gaaacagcta tgaccatgat tacgaattcg gcttgcggcc gcttctagag ggcgcgcctt
     2521 tacagctagc tcagtcctag gtattatgct agccagttag ctgtcaccgg atgtgctttc
     2581 cggtctgatg agtccgtgag gacgaaacag cctctacaaa taattttgtt taatcacaca
     2641 ggaaagtact agatgaaagt ggccatttat gttcgtgtta gcaccgatga acaggccaaa
     2701 gaaggtttta gcattccggc acagcgtgaa cgtctgcgtg cattttgtgc aagccagggt
     2761 tgggaaattg tgcaagaata tattgaagaa ggttggagcg caaaagatct ggatcgtccg
     2821 cagatgcagc gtctgctgaa agatatcaaa aaaggcaaca ttgatattgt gctggtgtat
     2881 cgtctggatc gcctgacccg tagcgttctg gatctgtatc tgctgctgca gacctttgaa
     2941 aaatacaatg tggcatttcg tagcgccacc gaagtttatg ataccagcac cgcaatgggt
     3001 cgtctgttta ttaccctggt tgcagcactg gcacagtggg aacgtgaaaa tctggcagaa
     3061 cgtgttaaat ttggtatcga gcagatgatc gatgaaggta aaaaaccggg tggtcatagc
     3121 ccgtatggtt acaaatttga taaagacttc aattgcacca ttattgagga agaagcagac
     3181 gttgttcgta tgatctatcg catgtattgt gatggttatg gctatcgtag cattgcagat
     3241 cgtctgaatg aactgatggt taaaccgcgt attgccaaag aatggaatca taatagcgtg
     3301 cgtgatatcc tgaccaacga tatctatatt ggcacctatc gttggggtga taaagttgtt
     3361 ccgaataatc atccgcctat tattagcgaa accctgttca aaaaagccca gaaagaaaaa
     3421 gaaaaacgtg gcgttgatcg taaacgcgtt ggtaaatttc tgtttaccgg tctgctgcag
     3481 tgtggtaatt gtggtggcca taaaatgcag ggccattttg ataaacgtga gcagaaaacc
     3541 tattaccgtt gtaccaaatg tcaccgcatt accaacgaaa aaaacattct ggaaccgctg
     3601 ctggatgaaa ttcagctgct gattaccagc aaagaatact ttatgagcaa attcagcgac
     3661 cgctatgatc agcaagaggt tgttgatgtt agcgcactga caaaagaact ggaaaaaatc
     3721 aaacgccaga aagagaaatg gtacgatctg tatatggatg atcgtaaccc gattccgaaa
     3781 gaagaactgt ttgccaaaat taacgaactg aacaaaaaag aagaagaaat ctatagcaag
     3841 ctgagcgaag tggaagaaga taaagaaccg gttgaagaga aatataaccg cctgagcaaa
     3901 atgatcgatt ttaaacagca gtttgagcag gccaacgact ttaccaaaaa agagctgctg
     3961 ttcagcatct tcgaaaagat tgtgatttat cgcgagaaag gcaagctgaa aaaaatcacc
     4021 ctggattacc ccctgaaata agtggttagg ctagctacat ttaatgacaa tgtgcaggtt
     4081 gtcatattgg tcatttattc cgtaactcgg gtcataagga gtggcgtatt tttgtttggt
     4141 ttaatccaat gcaagaacgg aaatggactc gatttactca tttgccttta ttaagtcgag
     4201 ctaaggtggt taacagtaca acaaagcaaa taaataaggc ggatcgtgtg attgagtttg
     4261 aagcatcgga tcttcaacga gccaaaataa tcgggatcct ctagagtcga cctgcaggca
     4321 tgcaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     4381 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     4441 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     4501 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     4561 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     4621 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4681 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
