LOCUS       sequence-370384-        9906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo gene amplification (HamAmp) of nerolidol synthetic module at
            RPL25 locus (15 copy). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9906)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 9906)
  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: pIT7A6
FEATURES             Location/Qualifiers
     source          1..9906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1031..1405)
                     /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"
     promoter        complement(4342..4783)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(4348..4371)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    4666..4783
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(6829..6846)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6829..6845)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6838..6860)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7054..7073)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7173..7195
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7233..7251)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7319..7423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7424..7492,7493..8284)
                     /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(7642..7661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8455..9043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8944..8963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9197..9214
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9331..9352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9367..9372,9373..9390,9391..9397)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9405..9421
                     /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     9410..9432
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9429..9445
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9429..9445
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcgacaaaca taaaaaagtt cttgccgtta agaagctgaa gttgcttctc gctataccag
       61 aggggattat tgtcgggagt catgatgcga catttacccg gttgagtaag gaaggggtgg
      121 aggaatcaga agagagataa acaaacatac acgtcaaaac agtatagttc aagtaacaca
      181 tatagacatg tacacgcgcg attacgtgtg tacgtgcaca tgttaatgtt atatacatat
      241 atagatatag atatataatt gcaataatga actacaaatt attatacccc cgttcttttt
      301 tttttctttt ttttagtaat ttttctttag atttatcgga atacttactt tggtgcttca
      361 ccttcggcgg cagcaggtgg ctgctgttgc tgatgttgtt gttgttgttg ttggtcttca
      421 gcttgaccgg actcggacat gtctgaagtc cataaggtta aattgtccct tagcagttgc
      481 atgataagtg tgctatcttt gtatgattct tcagacagag tgtccaactc agcaatagcg
      541 tcgtcaaaag cttgcttggc caaatggcag gctttgtctg gagagttttg aatttcgcat
      601 gcaaatcatt acgaccgaga ttcccgggac gccacagaaa cctcagaata agctaaaggg
      661 taatataata aagcccttcg attcaataat agcattaccc aagaggcata gcggcaaact
      721 aagttgacaa cacaaaatca aatcgcttca tttttatttt attaatgata ttttattaga
      781 gaggaaatta tgtaatatct agatattaaa tataaagtag atctctgtct aaaatattct
      841 taacaaaatc ttatattcga aaggatgacg gtccaatcaa aggattgtat aagagaaatt
      901 agttttcgaa atcaattttt acttgacctg aacttaacga tttgctctca atccgccatg
      961 gtcactatta tggacctggg ttaagttcaa cgtcaccagc caacttcaac aaagagaagt
     1021 tagtagcacc aactctttta acgaaaaccc agtaagagtc accagataaa gcctttttca
     1081 tgtgaacttt aaccttagtt ggacctctgg aagttgggat catccattca aacatagtgt
     1141 ttaagttacc agaagcaaca ccttccttaa acttaccata gaactctgga gagtcagtac
     1201 ctggagcaac gtccttaaag aagttcttac caataacttg ttttggatct ctaccagtaa
     1261 tatcaccttc agcagcattg tattgcaaaa tgttaccatc accgtccaat tgaatagcac
     1321 cgaaagctaa accgtctaat tgaccatcat ccattttagc caaagtgttt tcaatatctt
     1381 cagaaccgaa agcaacgtgt tccattttgg atccactgta aaaaactttt tttattatac
     1441 tattttctat tgtttaatag tcgttgttct tgtggatata tgaacaagta atcaaaaaag
     1501 ttatcacaaa tgaactcaga tatatttcag ccgaatggct agagtttata tatgtctgtc
     1561 taacccacac tttttccgca ttttattcct caactcatcc ctcccttgaa caacgactgc
     1621 taacaacgtc tcactctaaa atctttacac gtttagcttt acaggaggag cataattgac
     1681 ttttgttcct atttcaagtt gggaagggct tactttagca ttagctcgga gtggtccacc
     1741 ccggaactat ggcctgcgga aatactctca caggcacatg gcccctgaac tgcggggagt
     1801 gtgccgcacg gaagagtgag cgccggtgcc taccggaaga accccttcct cagtatttga
     1861 gcacgtgact cgaatgctgg ccgcaaaaaa gaaaaaagaa tgtaatccgg ttgagccgcc
     1921 ccggaagacg cttttccggt agttaattct ttatacatta ctggaacaga ttctgctccg
     1981 tttaattaga cttggatgca aggttgcctg tgatagcttt agtgggtcta ttctacggcc
     2041 ttctaatggc accgttatga cctggatctg gtacgtctgt acgattagcg gggcaagttc
     2101 aaataaaatt ggtttactcg agttaattta tctcatgaag taatgatacc tcagcgatgt
     2161 gctcgctcgt tagtatgctt cataaattca ttattatttt tttttggaag tgtttttcga
     2221 caaaagttgc atttttacga aggatatgta aactcaagaa gtactaaata aatataaata
     2281 caattcgaaa atttgaaaaa agtaggagac aacactttcc ttcctcacga agaagttaag
     2341 cttgagcatc ggaccgaagc ataagaatgc agcaaattga taatattccc gttaagccag
     2401 tcgtaggaaa aggaaaagaa tgagtagcaa atgtcacaca acaaacgcgc accatcccta
     2461 catacacaaa cacacacaca cacacacata tatatatatt ctaccccata taagcgagtg
     2521 attcatcatg aaaagaattt atcacttcga atttaaataa taacattctt acctgatgta
     2581 caactcttta ctctcatctt gcgaaaagaa aagggcaggg tgccgctgaa acagcggcgt
     2641 tgctgatgaa aataaccctg tgttttcgcc tcattctctc aaaaccttca cagatccttt
     2701 tcttgttata gcctcaataa aatcacagta cactaacacg taattataaa tagtaaccgg
     2761 aaccctcgct gcaatacgca aattggtacc tactttgtaa ggcacaggaa accccgtgca
     2821 ctctgccact acactttatt tacgcttaat catgaattta acatgttatg aaagatttat
     2881 tgtatattta aagaaaaatt taaaaataat attaaattta ttaattaaac caattagagc
     2941 taggggccca ttgtagcaaa gattgtaagg aaatagcaaa ccagaggtcc aagtcaaaac
     3001 ggcaaaaaag aagaaagtaa gcaatataat ggacgggttt acatgtttaa atatatgtac
     3061 atgtatgtag tcttatactt tcaaaacata atcgcatgcg ggtcatcaaa ataaacctcc
     3121 attttggact tgggaaaaag aaaatttgat ttaaactgtt gtatgtagct aacaatacat
     3181 aaaactattt aaataaaata ggaataagaa tgctcaatta aaaaatgatt tattttctaa
     3241 caaaacttcc tgcaggtcac tattatttgc ttctcttgta aactttgttc aaaaacgcag
     3301 ttaagacgtc ggctttgaag ccacgagact cgtcgacttg ggagatcttg gccttcaaat
     3361 ccttggcaac agactcttca tattcgtggt ataactggtc gattttcaag tcattgaaaa
     3421 tctttttgca tttggcttct gcgactgagt ccttcttacc gtaattttcg tctaaagtct
     3481 ttctttgttc tgcggaagca agttctaatg ccttgttgat tacccaagaa catttgttat
     3541 cttggatatc tgtaccgatc ttaccgatct gttctggggt accgaagcag tctaagtagt
     3601 catcttgaat ttggaaatat tcacccaatg gaatcaagac atctctggct tgtttcaaat
     3661 ccttttcatc tgtgataccg gcaacgtaca tagccaatgc gacaggcaag tagaaagaat
     3721 agtaagcagt cttgaaagta actatgaagg agtgcttctt tagggagaac ttactcaagt
     3781 cgactttgtc ttcaggtgca gtgattaagt ccatcaattg gcccaattcg gtttggaagg
     3841 tgacttcatg gaacaattcg gtgatatcta tgtagtattt ttcgtttctg aagtgagatt
     3901 tcaaaagctt gtagatagca gcctctaaca tgaatgcgtc attgatggca atttccccaa
     3961 cttcaggaac cttgtaccaa catggttggc ctcttctggt aatggacttg tccatcatat
     4021 catcggcgac caagaagtaa gcctgcaaca actcaatgca ccaacctaga atagcaacct
     4081 tttcgtattc ttcttgcccc aattgttcaa cggtcttgtt ggagagaata gcatacgtgt
     4141 ccacaacgga caaacctcta tttaacttac cgcctggagt gttgtagttc aatgagtggg
     4201 cataccagtc acatgcttcc ttaggcatac cgtaagccaa aagcgatgcg ttcaattcct
     4261 ctactaattt agggaaaacg ttcaagaatc tctctctcct aatttctttt tctgaagcca
     4321 ttttgctagc tatagttttt tctccttgac gttaaagtat agaggtatat taacaatttt
     4381 ttgttgatac ttttatgaca tttgaataag aagtaataca aaccgaaaat gttgaaagta
     4441 ttagttaaag tggttatgca gcttttgcat ttatatatct gttaatagat caaaaatcat
     4501 cgcttcgctg attaattacc ccagaaataa ggctaaaaaa ctaatcgcat tattatccta
     4561 tggttgttaa tttgattcgt tgatttgaag gtttgtgggg ccaggttact gccaattttt
     4621 cctcttcata accataaaag ctagtattgt agaatcttta ttgttcggag cagtgcggcg
     4681 cgaggcacat ctgcgtttca ggaacgcgac cggtgaagac caggacgcac ggaggagagt
     4741 cttccgtcgg agggctgtcg cccgctcggc ggcttctaat ccgtacttca atatagcaat
     4801 gagcagttaa gcgtattact gaaagttcca aagagaaggt ttttttaggc taagataatg
     4861 gggctcttta catttccaca acatataagt aagattagat atggatatgt atatggtggt
     4921 attgccatgt aatatgatta ttaaacttct ttgcgtccat ccaaaaaaaa agtaagaatt
     4981 tttgaaaatt caatataagc tagcgcaggc taaccggaac ctgtattatt tagtttatgc
     5041 tacgttaaat aaagaccttt cgttcacata actgaatgtg taatggcctt gagatttcaa
     5101 gcataccaag ttggtggaga cggggtcgtt acaaaagact ctttaagcca agatttcctt
     5161 gacagccttg gcgatagcat cgccaacctc agtggtagag ttagaaccac caaggtcacc
     5221 ggttctgaca cctgcatcca agacatttct aacagcttct tcaagagccc taccttcttc
     5281 aaccaaatcc aaggataact tcaacatcat agctgcagat aagatggtag caattgggtt
     5341 aaccttgttt gctggtaaat ctggggcaga accatgacat ggttcgtaca aaccgaatgc
     5401 cttgttagtg tcaggtaggg aagctagaga tgcagaaggt aataaaccca aagaacctgg
     5461 aataacagag gcttcatcgg agataatatc accaaacatg ttgttggtaa taacaacacc
     5521 gtttagctta gttggtgatt taaccaaaat catagcagca gagtcgatca attggtgctg
     5581 aacagttaat tgtgggaact cagtcttgat ggtttcttca acagtctttc tccacaatct
     5641 ggaagaggca agcacgttag ccttgtcaag agaccagatt ggtaatggtg ggttttgttg
     5701 caatgccaag aaagcagcca ttcttgtaat tctttgaact tcaggaacac tgtatttctc
     5761 agagtcccaa gcaactccgt caccttcatc ttcttttctt tcaccaaagt agataccacc
     5821 aaccaattct ctaacgacga cgaaatcggt accctttgca tattcaggct tcaaaggaga
     5881 aagatctagt aaagaatcag aagcaaagtt acatggtctc aagttggcgt atagacccaa
     5941 ttcctttctg atcttcaata gaccttgttc tggtctaact gcgcccgtac cccattttgg
     6001 accaccaaca gcacctagta agacagcatc ggctttctta gaggcttcta gagcttcatc
     6061 tggtaaagga gtgccagtgg catcgatggc agcacccccg atcaagtgat gttggaaatt
     6121 gaacttaatt tctggacgga cttcagcaat ggcattcaag accttaatag cttcgtcagt
     6181 aacttcttta ccgacgtgat cacccggtag gacaacgata ttcttagaca ttttgcggcc
     6241 gctttatctt attgatcttc tttgtttagc cttttcaaca gcagaagaaa agcatatgag
     6301 atacatttaa aattgtttta aaataatttt aaaagataat atctaaaaga ttgtatgcat
     6361 acgaacgaag ttgcattggg aagccccaac gcagtgaggc aagccgaagg tagaaagaaa
     6421 atccccttgt gcgtccagtg agaaacagca agcagtgggt aggaaggccc agtttagcct
     6481 gactgttcca acggggctcc caggctgagg aaatgcttct ctccgtaatg ggagagccta
     6541 actccagagc gtggacttcg gctcacggtt ttctcttttt ttggcgcaca atgactgctg
     6601 agacaaaaaa agaaagcaac tccatgtaaa cacccgtaca tattaaataa gtgacccatg
     6661 cctcctcatc tcttcattgc aaagtacgga aaccgttaga ttggtaccaa caaggttgct
     6721 atatcgggca tacaagataa gaaacttaaa gaacgccgag acaagaatat gtaatgataa
     6781 gcctcctccg gggtgttaga ctgattagta catgtttaaa ctgaattcac tggccgtcgt
     6841 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     6901 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     6961 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     7021 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7081 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     7141 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     7201 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     7261 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7321 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7381 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7441 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7501 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7561 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7621 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7681 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7741 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7801 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7861 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7921 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7981 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     8041 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8101 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     8161 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8221 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8281 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8341 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8401 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8461 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8521 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8581 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8641 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8701 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8761 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8821 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8881 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8941 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     9001 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     9061 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     9121 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     9181 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     9241 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     9301 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     9361 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     9421 atttcacaca ggaaacagct atgaccatga ttacgccaag cttgtttaaa ctatgttcct
     9481 tgatacctcg acgtggagca cgaaatagaa gaaaaaagta taatgaggat ggaataattt
     9541 tcttgaatct gagcagaaaa acgaacgaac actctaatgg atttgttaat ggctgttggt
     9601 agtaaaaaga acaacttcca cagcatcaac gcttctatca aaacgattgc taataaactg
     9661 ttgagcccga taattacgtt gctcaccata tcttaatcaa gtccttaatg cactattctt
     9721 gtcgtcggat agtttattta cgtgtggttt tcgtatctca tcaatatcaa ctctactagt
     9781 tcccatctat cttcctctca attttctcgt gctattaata ttgcagttca cataccagat
     9841 ggagccattg gcacaaatgt ggtttccttt cttattgctg tatccaactt ttgtgaagct
     9901 acaggt
//
