LOCUS       sequence-370865-        7555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo gene amplification (HamAmp) of EforRed expression module at 
            RPL25 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7555)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 7555)
  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: pIEforRed
FEATURES             Location/Qualifiers
     source          1..7555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1222..1492
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(2279..2295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2279..2295)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2292..2314)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2303..2319
                     /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(2327..2333,2334..2351,2352..2357))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2372..2393
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2510..2527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2681..3269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2761..2780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3440..4231,4232..4300))
                     /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     4063..4082
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4301..4405)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4473..4491
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4529..4551)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4651..4670
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4864..4886
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4878..4895
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4879..4895
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcatatggcg tatccaagcc gaaacggcgc tcgcctcatc cccacgggaa taaggcagcc
       61 gacaaaagaa aaacgaccga aaaggaacca gaaagaaaaa agagggtggg cgcgccgcgg
      121 acgtgtaaaa agatatgcat ccagcttcta tatcgcttta actttaccgt tttgggcatc
      181 gggaacgtat gtaacattga tctcctcttg ggaacggtga gtgcaacgaa tgcgatatag
      241 caccgaccat gtgggcaaat tcgtaataaa ttcggggtga gggggattca agacaagcaa
      301 ccttgttagt cagctcaaac agcgatttaa cggttgagta acacatcaaa acaccgttcg
      361 aggtcaagcc tggcgtgttt aacaagttct tgatatcata tataaatgta ataagaagtt
      421 tggtaatatt caattcgaag tgttcagtct tttacttctc ttgttttata gaagaaaaaa
      481 catcaagaaa catctttaac atacacaaac acatactatc agaatacagg atccaaaatg
      541 tctgttatca agcaagttat gaagactaaa ttgcatttgg aaggtactgt taacggtcat
      601 gatttcacta ttgaaggtaa aggtgagggt aaaccatacg aaggtttaca acacatgaag
      661 atgactgtca ctaaaggtgc tccattgcca ttttccgttc atattttgac tccatctcac
      721 atgtatggtt ctaagccatt taacaaatac ccagctgaca ttcctgatta ccacaaacaa
      781 tcttttccag aaggtatgtc ttgggagaga tccatgattt ttgaagatgg tggtgtttgt
      841 accgcttcta atcattcttc tatcaacttg caagagaatt gcttcatcta cgatgttaag
      901 tttcacggtg ttaacttgcc accagatggt ccagttatgc aaaagactat tgctggttgg
      961 gaaccatctg ttgaaacttt gtacgttaga gatggtatgt tgaaatctga taccgctatg
     1021 gttttcaagt tgaaaggtgg tggtcatcac agagttgatt tcaaaactac ttacaaggct
     1081 aagaaaccag ttaagttgcc agaatttcat ttcgtcgagc atagattgga attgaccaaa
     1141 cacgataagg atttcactac ttgggatcaa caagaagctg ctgaaggtca tttttcacct
     1201 ttgccaaagg ctttgccata aattgaattg aattgaaatc gatagatcaa tttttttctt
     1261 ttctctttcc ccatccttta cgctaaaata atagtttatt ttattttttg aatatttttt
     1321 atttatatac gtatatatag actattattt atcttttaat gattattaag atttttatta
     1381 aaaaaaaatt cgctcctctt ttaatgcctt tatgcagttt ttttttccca ttcgatattt
     1441 ctatgttcgg gttcagcgta ttttaagttt aataactcga aaattctgcg ttcgttaaag
     1501 ctttcgagaa ggatattatt tcccgggaat ctcggtcgta atgatttgca tgcccgccat
     1561 ctctactcac tccataatat tacatataga tataggacaa gcccgcattt tcatactgaa
     1621 aggtaaactt ctattattat agtggtatcc aacgttcacc gcttccagca tagcagaaat
     1681 tacgtgtttt tgcatatgtt atgctgatca ttgtatgctt actaccattt ttctttgctt
     1741 cgccttgcct tctttgacgt ttttttgaag caaaaaaaaa gtcaagacag atgtgcttac
     1801 aaaaccatgt aaggctcatt ttcaaagaag ctactaatag aaagagaaca aagcgtttac
     1861 gagtctggaa aatcaatggc tccatctggt atgtgaactg caatattaat agcacgagaa
     1921 aattgagagg aagatagatg ggaactagta gagttgatat tgatgagata cgaaaaccac
     1981 acgtaaataa actatccgac gacaagaata gtgcattaag gacttgatta agatatggtg
     2041 agcaacgtaa ttatcgggct caacagttta ttagcaatcg ttttgataga agcgttgatg
     2101 ctgtggaagt tgttcttttt actaccaaca gccattaaca aatccattag agtgttcgtt
     2161 cgtttttctg ctcagattca agaaaattat tccatcctca ttatactttt ttcttctatt
     2221 tcgtgctcca cgtcgaggta tcaaggaaca tagtttaaac aagcttggcg taatcatggt
     2281 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2341 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2401 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3121 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3541 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3601 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3661 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3721 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3781 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3841 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3901 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3961 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4021 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4081 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4141 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4201 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4261 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4381 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     4441 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     4501 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4561 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     4621 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     4681 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     4741 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4801 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     4861 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cagtttaaac atgtactaat
     4921 cagtctaaca ccccggagga ggcttatcat tacatattct tgtctcggcg ttctttaagt
     4981 ttcttatctt gtatgcccga tatagcaacc ttgttggtac caatctaacg gtttccgtac
     5041 tttgcaatga agagatgagg aggcatgggt cacttattta atatgtacgg gtgtttacat
     5101 ggagttgctt tctttttttg tctcagcagt cattgtgcgc caaaaaaaga gaaaaccgtg
     5161 agccgaagtc cacgctctgg agttaggctc tcccattacg gagagaagca tttcctcagc
     5221 ctgggagccc cgttggaaca gtcaggctaa actgggcctt cctacccact gcttgctgtt
     5281 tctcactgga cgcacaaggg gattttcttt ctaccttcgg cttgcctcac tgcgttgggg
     5341 cttcccaatg caacttcgtt cgtatgcata caatctttta gatattatct tttaaaatta
     5401 ttttaaaaca attttaaatg tatctcatat gcttttcttc tgctgttgaa aaggctaaac
     5461 aaagaagatc aataagataa agcggccgca aaatgtctaa gaatatcgtt gtcctaccgg
     5521 gtgatcacgt cggtaaagaa gttactgacg aagctattaa ggtcttgaat gccattgctg
     5581 aagtccgtcc agaaattaag ttcaatttcc aacatcactt gatcgggggt gctgccatcg
     5641 atgccactgg cactccttta ccagatgaag ctctagaagc ctctaagaaa gccgatgctg
     5701 tcttactagg tgctgttggt ggtccaaaat ggggtacggg cgcagttaga ccagaacaag
     5761 gtctattgaa gatcagaaag gaattgggtc tatacgccaa cttgagacca tgtaactttg
     5821 cttctgattc tttactagat ctttctcctt tgaagcctga atatgcaaag ggtaccgatt
     5881 tcgtcgtcgt tagagaattg gttggtggta tctactttgg tgaaagaaaa gaagatgaag
     5941 gtgacggagt tgcttgggac tctgagaaat acagtgttcc tgaagttcaa agaattacaa
     6001 gaatggctgc tttcttggca ttgcaacaaa acccaccatt accaatctgg tctcttgaca
     6061 aggctaacgt gcttgcctct tccagattgt ggagaaagac tgttgaagaa accatcaaga
     6121 ctgagttccc acaattaact gttcagcacc aattgatcga ctctgctgct atgattttgg
     6181 ttaaatcacc aactaagcta aacggtgttg ttattaccaa caacatgttt ggtgatatta
     6241 tctccgatga agcctctgtt attccaggtt ctttgggttt attaccttct gcatctctag
     6301 cttccctacc tgacactaac aaggcattcg gtttgtacga accatgtcat ggttctgccc
     6361 cagatttacc agcaaacaag gttaacccaa ttgctaccat cttatctgca gctatgatgt
     6421 tgaagttatc cttggatttg gttgaagaag gtagggctct tgaagaagct gttagaaatg
     6481 tcttggatgc aggtgtcaga accggtgacc ttggtggttc taactctacc actgaggttg
     6541 gcgatgctat cgccaaggct gtcaaggaaa tcttggctta aagagtcttt tgtaacgacc
     6601 ccgtctccac caacttggta tgcttgaaat ctcaaggcca ttacacattc agttatgtga
     6661 acgaaaggtc tttatttaac gtagcataaa ctaaataata caggttccgg ttagcctgcg
     6721 ggcccaaagc tagctctaat tggtttaatt aataaattta atattatttt taaatttttc
     6781 tttaaatata caataaatct ttcataacat gttaaattca tgattaagcg taaataaagt
     6841 gtagtggcag agtgcacggg gtttcctgtg ccttacaaag taggtaccaa tttgcgtatt
     6901 gcagcgaggg ttccggttac tatttataat tacgtgttag tgtactgtga ttttattgag
     6961 gctataacaa gaaaaggatc tgtgaaggtt ttgagagaat gaggcgaaaa cacagggtta
     7021 ttttcatcag caacgccgct gtttcagcgg caccctgccc ttttcttttc gcaagatgag
     7081 agtaaagagt tgtacatcag gtaagaatgt tattatttaa attcgaagtg ataaattctt
     7141 ttcatgatga atcactcgct tatatggggt agaatatata tatatgtgtg tgtgtgtgtg
     7201 tgtttgtgta tgtagggatg gtgcgcgttt gttgtgtgac atttgctact cattcttttc
     7261 cttttcctac gactggctta acgggaatat tatcaatttg ctgcattctt atgcttcggt
     7321 ccgatgctca agcttaactt cttcgtgagg aaggaaagtg ttgtctccta cttttttcaa
     7381 attttcgaat tgtatttata tttatttagt acttcttgag tttacatatc cttcgtaaaa
     7441 atgcaacttt tgtcgaaaaa cacttccaaa aaaaaataat aatgaattta tgaagcatac
     7501 taacgagcga gcacatcgct gaggtatcat tacttcatga gataaattaa ctcga
//
