LOCUS       sequence-149381-        2911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Staging plasmid for use in DIP transposition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2911)
  AUTHORS   Nadler DC, Morgan SA, Flamholz A, Kortright KE, Savage DF
  TITLE     Rapid construction of metabolite biosensors using domain-insertion 
            profiling.
  JOURNAL   Nat Commun. 2016 Jul 29;7:12266. doi: 10.1038/ncomms12266.
  PUBMED    27470466
REFERENCE   2  (bases 1 to 2911)
  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: "Nat 
            Commun."; date: "2016-07-29"; volume: "7:12266. doi"; pages: " 
            10.1038/ncomms12266"
COMMENT     Sequence Label: pATT-Dest
FEATURES             Location/Qualifiers
     source          1..2911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..148
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(149..217,218..1009)
                     /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     complement(367..386)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1180..1768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1669..1688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1922..1939
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2038..2057
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2038..2056
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2088..2110
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2198..2219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2234..2239,2240..2257,2258..2264)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2272..2288
                     /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     2277..2299
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2296..2312
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2296..2312
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2308..2631
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2325..2381
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2382..2399)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2382..2398)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2391..2413)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2607..2626)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2788..2806)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2802..2849
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 caaggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
       61 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      121 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      181 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      241 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      301 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      361 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      421 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      481 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      541 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      601 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      661 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      721 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      781 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      841 taaatctgga gccggtgagc gtggctctcg cggtatcatt gcagcactgg ggccagatgg
      901 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
      961 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1021 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1081 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1141 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1201 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1261 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1321 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1381 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1441 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1501 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1561 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1621 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1681 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1741 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1801 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1861 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1921 cagcgagtca gtgagcgagg aagcggaaga gcgccatcag caaccgcacc tgtggcgccg
     1981 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     2041 tacgactcac tatagggaca ccacaacggt ttccctctag aaataatttt gtttaacttt
     2101 aagaaggaga tataccatcg tctctgtcca gagaccatta atgcagctgg cacgacaggt
     2161 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     2221 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     2281 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc
     2341 aggtcgactc tagaggatcc ccgggtaccg agctcgaatt cactggccgt cgttttacaa
     2401 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2461 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2521 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2581 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2641 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     2701 gcttacagac aagctggtct ctagcgagag acggttaaga tccggctgct aacaaagccc
     2761 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     2821 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac
     2881 aggacgggtg tggtcgccat gatcgcgtag t
//
