LOCUS       sequence-370833-       10053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A backbone plasmid for in vivo gene amplification (HamAmp) of a 
            monoterpene synthetic module (+ GPP synthase without a monoterpene 
            synthase) at RPL25 locus (14 copy).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10053)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 10053)
  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"
COMMENT     Sequence Label: pIT6EG7m
FEATURES             Location/Qualifiers
     source          1..10053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1105..1121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1105..1121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1118..1140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1129..1145
                     /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)."
     promoter        complement(join(1153..1159,1160..1177,1178..1183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1198..1219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1336..1353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1507..2095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1587..1606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2266..3057,3058..3126))
                     /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     2889..2908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3127..3231)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3299..3317
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3355..3377)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3477..3496
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3690..3712
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3704..3721
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3705..3721
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             7093..7467
                     /codon_start=1
                     /product="small protein tag, derived from photoactive 
                     yellow protein, that allows in vivo fluorescent labeling by
                     addition of a cell-permeant fluorogen (Plamont et al., 
                     2016)"
                     /label=Y-FAST
                     /translation="MEHVAFGSEDIENTLAKMDDGQLDGLAFGAIQLDGDGNILQYNAA
                     EGDITGRDPKQVIGKNFFKDVAPGTDSPEFYGKFKEGVASGNLNTMFEWMIPTSRGPTK
                     VKVHMKKALSGDSYWVFVKRV"
     CDS             complement(8657..8674)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gtttctgtgg cgtgacccgg gaatctcggt cgtaatgatt tgcatgcgaa attcaaaact
       61 ctccagacaa agcctgccat ttggccaagc aagcttttga cgacgctatt gctgagttgg
      121 acactctgtc tgaagaatca tacaaagata gcacacttat catgcaactg ctaagggaca
      181 atttaacctt atggacttca gacatgtccg agtccggtca agctgaagac caacaacaac
      241 aacaacaaca tcagcaacag cagccacctg ctgccgccga aggtgaagca ccaaagtaag
      301 tattccgata aatctaaaga aaaattacta aaaaaaagaa aaaaaaaaga acgggggtat
      361 aataatttgt agttcattat tgcaattata tatctatatc tatatatgta tataacatta
      421 acatgtgcac gtacacacgt aatcgcgcgt gtacatgtct atatgtgtta cttgaactat
      481 actgttttga cgtgtatgtt tgtttatctc tcttctgatt cctccacccc ttccttactc
      541 aaccgggtaa atgtcgcatc atgactcccg acaataatcc cctctggtat agcgagaagc
      601 aacttcagct tcttaacggc aagaactttt ttatgtttgt cgcacctgta gcttcacaaa
      661 agttggatac agcaataaga aaggaaacca catttgtgcc aatggctcca tctggtatgt
      721 gaactgcaat attaatagca cgagaaaatt gagaggaaga tagatgggaa ctagtagagt
      781 tgatattgat gagatacgaa aaccacacgt aaataaacta tccgacgaca agaatagtgc
      841 attaaggact tgattaagat atggtgagca acgtaattat cgggctcaac agtttattag
      901 caatcgtttt gatagaagcg ttgatgctgt ggaagttgtt ctttttacta ccaacagcca
      961 ttaacaaatc cattagagtg ttcgttcgtt tttctgctca gattcaagaa aattattcca
     1021 tcctcattat acttttttct tctatttcgt gctccacgtc gaggtatcaa ggaacatagt
     1081 ttaaacaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1141 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1201 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1261 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1321 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1381 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1441 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1501 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1561 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1621 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1681 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1741 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1801 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1861 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1921 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1981 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2041 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2101 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2161 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2221 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2281 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2341 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2401 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2461 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2521 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2581 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2641 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2701 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2761 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2821 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2881 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2941 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3001 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3061 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3121 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3181 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3241 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3301 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     3361 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3421 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     3481 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     3541 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     3601 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     3661 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     3721 tgaattcagt ttaaacatgt actaatcagt ctaacacccc ggaggaggct tatcattaca
     3781 tattcttgtc tcggcgttct ttaagtttct tatcttgtat gcccgatata gcaaccttgt
     3841 tggtaccaat ctaacggttt ccgtactttg caatgaagag atgaggaggc atgggtcact
     3901 tatttaatat gtacgggtgt ttacatggag ttgctttctt tttttgtctc agcagtcatt
     3961 gtgcgccaaa aaaagagaaa accgtgagcc gaagtccacg ctctggagtt aggctctccc
     4021 attacggaga gaagcatttc ctcagcctgg gagccccgtt ggaacagtca ggctaaactg
     4081 ggccttccta cccactgctt gctgtttctc actggacgca caaggggatt ttctttctac
     4141 cttcggcttg cctcactgcg ttggggcttc ccaatgcaac ttcgttcgta tgcatacaat
     4201 cttttagata ttatctttta aaattatttt aaaacaattt taaatgtatc tcatatgctt
     4261 ttcttctgct gttgaaaagg ctaaacaaag aagatcaata agataaagcg gccgcaaaat
     4321 gtctaagaat atcgttgtcc taccgggtga tcacgtcggt aaagaagtta ctgacgaagc
     4381 tattaaggtc ttgaatgcca ttgctgaagt ccgtccagaa attaagttca atttccaaca
     4441 tcacttgatc gggggtgctg ccatcgatgc cactggcact cctttaccag atgaagctct
     4501 agaagcctct aagaaagccg atgctgtctt actaggtgct gttggtggtc caaaatgggg
     4561 tacgggcgca gttagaccag aacaaggtct attgaagatc agaaaggaat tgggtctata
     4621 cgccaacttg agaccatgta actttgcttc tgattcttta ctagatcttt ctcctttgaa
     4681 gcctgaatat gcaaagggta ccgatttcgt cgtcgttaga gaattggttg gtggtatcta
     4741 ctttggtgaa agaaaagaag atgaaggtga cggagttgct tgggactctg agaaatacag
     4801 tgttcctgaa gttcaaagaa ttacaagaat ggctgctttc ttggcattgc aacaaaaccc
     4861 accattacca atctggtctc ttgacaaggc taacgtgctt gcctcttcca gattgtggag
     4921 aaagactgtt gaagaaacca tcaagactga gttcccacaa ttaactgttc agcaccaatt
     4981 gatcgactct gctgctatga ttttggttaa atcaccaact aagctaaacg gtgttgttat
     5041 taccaacaac atgtttggtg atattatctc cgatgaagcc tctgttattc caggttcttt
     5101 gggtttatta ccttctgcat ctctagcttc cctacctgac actaacaagg cattcggttt
     5161 gtacgaacca tgtcatggtt ctgccccaga tttaccagca aacaaggtta acccaattgc
     5221 taccatctta tctgcagcta tgatgttgaa gttatccttg gatttggttg aagaaggtag
     5281 ggctcttgaa gaagctgtta gaaatgtctt ggatgcaggt gtcagaaccg gtgaccttgg
     5341 tggttctaac tctaccactg aggttggcga tgctatcgcc aaggctgtca aggaaatctt
     5401 ggcttaaaga gtcttttgta acgaccccgt ctccaccaac ttggtatgct tgaaatctca
     5461 aggccattac acattcagtt atgtgaacga aaggtcttta tttaacgtag cataaactaa
     5521 ataatacagg ttccggttag cctgcgggcc caaagctagc tctaattggt ttaattaata
     5581 aatttaatat tatttttaaa tttttcttta aatatacaat aaatctttca taacatgtta
     5641 aattcatgat taagcgtaaa taaagtgtag tggcagagtg cacggggttt cctgtgcctt
     5701 acaaagtagg taccaatttg cgtattgcag cgagggttcc ggttactatt tataattacg
     5761 tgttagtgta ctgtgatttt attgaggcta taacaagaaa aggatctgtg aaggttttga
     5821 gagaatgagg cgaaaacaca gggttatttt catcagcaac gccgctgttt cagcggcacc
     5881 ctgccctttt cttttcgcaa gatgagagta aagagttgta catcaggtaa gaatgttatt
     5941 atttaaattc gaagtgataa attcttttca tgatgaatca ctcgcttata tggggtagaa
     6001 tatatatata tgtgtgtgtg tgtgtgtgtt tgtgtatgta gggatggtgc gcgtttgttg
     6061 tgtgacattt gctactcatt cttttccttt tcctacgact ggcttaacgg gaatattatc
     6121 aatttgctgc attcttatgc ttcggtccga tgctcaagct taacttcttc gtgaggaagg
     6181 aaagtgttgt ctcctacttt tttcaaattt tcgaattgta tttatattta tttagtactt
     6241 cttgagttta catatccttc gtaaaaatgc aacttttgtc gaaaaacact tccaaaaaaa
     6301 aataataatg aatttatgaa gcatactaac gagcgagcac atcgctgagg tatcattact
     6361 tcatgagata aattaactcg agtaaaccaa ttttatttga acttgccccg ctaatcgtac
     6421 agacgtacca gatccaggtc ataacggtgc cattagaagg ccgtagaata gacccactaa
     6481 agctatcaca ggcaaccttg catccaagtc taattaaacg gagcagaatc tgttccagta
     6541 atgtataaag aattaactac cggaaaagcg tcttccgggg cggctcaacc ggattacatt
     6601 cttttttctt ttttgcggcc agcattcgag tcacgtgctc aaatactgag gaaggggttc
     6661 ttccggtagg caccggcgct cactcttccg tgcggcacac tccccgcagt tcaggggcca
     6721 tgtgcctgtg agagtatttc cgcaggccat agttccgggg tggaccactc cgagctaatg
     6781 ctaaagtaag cccttcccaa cttgaaatag gaacaaaagt caattatgct cctcctgtaa
     6841 agctaaacgt gtaaagattt tagagtgaga cgttgttagc agtcgttgtt caagggaggg
     6901 atgagttgag gaataaaatg cggaaaaagt gtgggttaga cagacatata taaactctag
     6961 ccattcggct gaaatatatc tgagttcatt tgtgataact tttttgatta cttgttcata
     7021 tatccacaag aacaacgact attaaacaat agaaaatagt ataataaaaa aagtttttac
     7081 agtggatcca aaatggaaca cgttgctttc ggttctgaag atattgaaaa cactttggct
     7141 aaaatggatg atggtcaatt agacggttta gctttcggtg ctattcaatt ggacggtgat
     7201 ggtaacattt tgcaatacaa tgctgctgaa ggtgatatta ctggtagaga tccaaaacaa
     7261 gttattggta agaacttctt taaggacgtt gctccaggta ctgactctcc agagttctat
     7321 ggtaagttta aggaaggtgt tgcttctggt aacttaaaca ctatgtttga atggatgatc
     7381 ccaacttcca gaggtccaac taaggttaaa gttcacatga aaaaggcttt atctggtgac
     7441 tcttactggg ttttcgttaa aagagttggt gctactaact tctctttgtt gaagttggct
     7501 ggtgacgttg aacttaaccc aggtccaaaa attgaagaag gtaagttggt tatctggatt
     7561 aacggtgaca aaggttacaa cggtttagcc gaagtcggta agaagtttga aaaggatact
     7621 ggtatcaaag tcaccgtcga acacccagac aaattagaag aaaaattccc acaagttgct
     7681 gctactggtg acggtccaga tattatcttt tgggctcacg atagatttgg tggttacgcc
     7741 caatctggtt tgttagctga aattacccca gataaggctt tccaagataa gttgtaccca
     7801 ttcacttggg atgctgtcag atacaacggt aagttgattg cttacccaat tgctgttgaa
     7861 gccttgtctt tgatctataa caaggatttg ttacctaatc caccaaaaac ttgggaagaa
     7921 atcccagctt tggataaaga attgaaagct aaaggtaaat ccgctttgat gttcaactta
     7981 caagaaccat acttcacttg gcctttaatc gctgctgacg gtggttacgc ttttaagtac
     8041 gaaaatggta aatacgatat taaggacgtt ggtgttgaca acgccggtgc taaagctggt
     8101 ttgactttct tggtcgattt gattaaaaac aagcatatga atgctgacac cgattactcc
     8161 atcgctgaag ctgctttcaa caaaggtgaa actgctatga ccatcaacgg tccttgggcc
     8221 tggtccaaca ttgacacttc taaggtcaac tatggtgtta ctgtcttgcc tacctttaag
     8281 ggtcaaccat ccaaaccatt tgttggtgtt ttgtctgctg gtatcaacgc tgcttctcca
     8341 aataaggaat tggctaagga atttttggaa aactacttgt taactgacga aggtttggaa
     8401 gccgttaaca aagataaacc attgggtgct gtcgccttga aatcttacga agaagaattg
     8461 gctaaagatc caagaattgc tgctactatg gaaaacgctc aaaagggtga aatcatgcct
     8521 aatattccac aaatgtctgc tttctggtat gccgtccgta ctgctgtcat caacgccgct
     8581 tccggtagac aaactgttga cgaagccttg aaggatgccc aaactagaat cactaagcca
     8641 ggttctggtg agctcggtgg tggtggtggt ggtgcttcag aaaaagaaat taggagagag
     8701 agattcttga acgttttccc taaattagta gaggaattga acgcatcgct tttggcttac
     8761 ggtatgccta aggaagcatg tgactggtat gcccactcat tgaactacaa cactccaggc
     8821 ggtaagctaa atagaggttt gtccgttgtg gacacgtatg ctattctctc caacaagacc
     8881 gttgaacaat tggggcaaga agaatacgaa aaggttgcca ttctaggttg gtgcattgag
     8941 ttgttgcagg cttactggtt ggtcgccgat gatatgatgg acaagtccat taccagaaga
     9001 ggccaaccat gttggtacaa ggttcctgaa gttggggaaa ttgccatctg ggacgcattc
     9061 atgttagagg ctgctatcta caagcttttg aaatctcact tcagaaacga aaaatactac
     9121 atagatatca ccgaattgtt ccatgaggtc accttccaaa ccgaattggg ccaattgatg
     9181 gacttaatca ctgcacctga agacaaagtc gacttgagta agttctccct aaagaagcac
     9241 tccttcatag ttactttcaa gactgcttac tattctttct acttgcctgt cgcattggcc
     9301 atgtacgttg ccggtatcac ggatgaaaag gatttgaaac aagccagaga tgtcttgatt
     9361 ccattgggtg aatacttcca aattcaagat gactacttag actgcttcgg taccccagaa
     9421 cagatcggta agatcggtac agatatccaa gataacaaat gttcttgggt aatcaacaag
     9481 gcattagaac ttgcttccgc agaacaaaga aagactttag acgaaaatta cggtaagaag
     9541 gactcagtcg cagaagccaa atgcaaaaag attttcaatg acttgaaaat tgaacagcta
     9601 taccacgaat atgaagagtc tattgccaag gatttgaagg ccaaaatttc tcaggtcgat
     9661 gagtctcgtg gcttcaaagc tgatgtctta actgcgttct tgaacaaggt ttacaagaga
     9721 agtaaatagt aaccatgggc ggattgagag caaatcgtta agttcaggtc aagtaaaaat
     9781 tgatttcgaa aactaatttc tcttatacaa tcctttgatt ggaccgtcat cctttcgaat
     9841 ataagatttt gttaagaata ttttagacag agatctactt tatatttaat atctagatat
     9901 tacataattt cctctctaat aaaatatcat taataaaata aaaatgaagc gatttgattt
     9961 tgtgttgtca acttagtttg ccgctatgcc tcttgggtaa tgctattatt gaatcgaagg
    10021 gctttattat attacccttt agcttattct gag
//
