LOCUS       sequence-370863-        7567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo gene amplification (HamAmp) of AeBlue expression module at 
            RPL25 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7567)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 7567)
  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: pIAeBlue
FEATURES             Location/Qualifiers
     source          1..7567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(784..800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(784..800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(797..819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    808..824
                     /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(832..838,839..856,857..862))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1015..1032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1186..1774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1266..1285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1945..2736,2737..2805))
                     /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     2568..2587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2806..2910)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2978..2996
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3034..3056)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3156..3175
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3369..3391
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3383..3400
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3384..3400
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      7294..7564
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
ORIGIN
        1 taaagctttc gagaaggata ttatttcccg ggaatctcgg tcgtaatgat ttgcatgccc
       61 gccatctcta ctcactccat aatattacat atagatatag gacaagcccg cattttcata
      121 ctgaaaggta aacttctatt attatagtgg tatccaacgt tcaccgcttc cagcatagca
      181 gaaattacgt gtttttgcat atgttatgct gatcattgta tgcttactac catttttctt
      241 tgcttcgcct tgccttcttt gacgtttttt tgaagcaaaa aaaaagtcaa gacagatgtg
      301 cttacaaaac catgtaaggc tcattttcaa agaagctact aatagaaaga gaacaaagcg
      361 tttacgagtc tggaaaatca atggctccat ctggtatgtg aactgcaata ttaatagcac
      421 gagaaaattg agaggaagat agatgggaac tagtagagtt gatattgatg agatacgaaa
      481 accacacgta aataaactat ccgacgacaa gaatagtgca ttaaggactt gattaagata
      541 tggtgagcaa cgtaattatc gggctcaaca gtttattagc aatcgttttg atagaagcgt
      601 tgatgctgtg gaagttgttc tttttactac caacagccat taacaaatcc attagagtgt
      661 tcgttcgttt ttctgctcag attcaagaaa attattccat cctcattata cttttttctt
      721 ctatttcgtg ctccacgtcg aggtatcaag gaacatagtt taaacaagct tggcgtaatc
      781 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      841 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      901 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      961 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1021 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1081 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1141 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1201 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1261 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1321 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1381 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1441 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1501 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1561 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1621 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1681 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1741 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1801 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1861 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1921 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1981 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2041 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2101 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2161 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2221 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2281 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2341 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2401 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2461 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2521 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2581 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2641 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2701 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2761 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2821 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2881 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2941 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     3001 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     3061 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     3121 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     3181 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3241 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3301 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     3361 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcagtt taaacatgta
     3421 ctaatcagtc taacaccccg gaggaggctt atcattacat attcttgtct cggcgttctt
     3481 taagtttctt atcttgtatg cccgatatag caaccttgtt ggtaccaatc taacggtttc
     3541 cgtactttgc aatgaagaga tgaggaggca tgggtcactt atttaatatg tacgggtgtt
     3601 tacatggagt tgctttcttt ttttgtctca gcagtcattg tgcgccaaaa aaagagaaaa
     3661 ccgtgagccg aagtccacgc tctggagtta ggctctccca ttacggagag aagcatttcc
     3721 tcagcctggg agccccgttg gaacagtcag gctaaactgg gccttcctac ccactgcttg
     3781 ctgtttctca ctggacgcac aaggggattt tctttctacc ttcggcttgc ctcactgcgt
     3841 tggggcttcc caatgcaact tcgttcgtat gcatacaatc ttttagatat tatcttttaa
     3901 aattatttta aaacaatttt aaatgtatct catatgcttt tcttctgctg ttgaaaaggc
     3961 taaacaaaga agatcaataa gataaagcgg ccgcaaaatg tctaagaata tcgttgtcct
     4021 accgggtgat cacgtcggta aagaagttac tgacgaagct attaaggtct tgaatgccat
     4081 tgctgaagtc cgtccagaaa ttaagttcaa tttccaacat cacttgatcg ggggtgctgc
     4141 catcgatgcc actggcactc ctttaccaga tgaagctcta gaagcctcta agaaagccga
     4201 tgctgtctta ctaggtgctg ttggtggtcc aaaatggggt acgggcgcag ttagaccaga
     4261 acaaggtcta ttgaagatca gaaaggaatt gggtctatac gccaacttga gaccatgtaa
     4321 ctttgcttct gattctttac tagatctttc tcctttgaag cctgaatatg caaagggtac
     4381 cgatttcgtc gtcgttagag aattggttgg tggtatctac tttggtgaaa gaaaagaaga
     4441 tgaaggtgac ggagttgctt gggactctga gaaatacagt gttcctgaag ttcaaagaat
     4501 tacaagaatg gctgctttct tggcattgca acaaaaccca ccattaccaa tctggtctct
     4561 tgacaaggct aacgtgcttg cctcttccag attgtggaga aagactgttg aagaaaccat
     4621 caagactgag ttcccacaat taactgttca gcaccaattg atcgactctg ctgctatgat
     4681 tttggttaaa tcaccaacta agctaaacgg tgttgttatt accaacaaca tgtttggtga
     4741 tattatctcc gatgaagcct ctgttattcc aggttctttg ggtttattac cttctgcatc
     4801 tctagcttcc ctacctgaca ctaacaaggc attcggtttg tacgaaccat gtcatggttc
     4861 tgccccagat ttaccagcaa acaaggttaa cccaattgct accatcttat ctgcagctat
     4921 gatgttgaag ttatccttgg atttggttga agaaggtagg gctcttgaag aagctgttag
     4981 aaatgtcttg gatgcaggtg tcagaaccgg tgaccttggt ggttctaact ctaccactga
     5041 ggttggcgat gctatcgcca aggctgtcaa ggaaatcttg gcttaaagag tcttttgtaa
     5101 cgaccccgtc tccaccaact tggtatgctt gaaatctcaa ggccattaca cattcagtta
     5161 tgtgaacgaa aggtctttat ttaacgtagc ataaactaaa taatacaggt tccggttagc
     5221 ctgcgggccc aaagctagct ctaattggtt taattaataa atttaatatt atttttaaat
     5281 ttttctttaa atatacaata aatctttcat aacatgttaa attcatgatt aagcgtaaat
     5341 aaagtgtagt ggcagagtgc acggggtttc ctgtgcctta caaagtaggt accaatttgc
     5401 gtattgcagc gagggttccg gttactattt ataattacgt gttagtgtac tgtgatttta
     5461 ttgaggctat aacaagaaaa ggatctgtga aggttttgag agaatgaggc gaaaacacag
     5521 ggttattttc atcagcaacg ccgctgtttc agcggcaccc tgcccttttc ttttcgcaag
     5581 atgagagtaa agagttgtac atcaggtaag aatgttatta tttaaattcg aagtgataaa
     5641 ttcttttcat gatgaatcac tcgcttatat ggggtagaat atatatatat gtgtgtgtgt
     5701 gtgtgtgttt gtgtatgtag ggatggtgcg cgtttgttgt gtgacatttg ctactcattc
     5761 ttttcctttt cctacgactg gcttaacggg aatattatca atttgctgca ttcttatgct
     5821 tcggtccgat gctcaagctt aacttcttcg tgaggaagga aagtgttgtc tcctactttt
     5881 ttcaaatttt cgaattgtat ttatatttat ttagtacttc ttgagtttac atatccttcg
     5941 taaaaatgca acttttgtcg aaaaacactt ccaaaaaaaa ataataatga atttatgaag
     6001 catactaacg agcgagcaca tcgctgaggt atcattactt catgagataa attaactcga
     6061 gcatatggcg tatccaagcc gaaacggcgc tcgcctcatc cccacgggaa taaggcagcc
     6121 gacaaaagaa aaacgaccga aaaggaacca gaaagaaaaa agagggtggg cgcgccgcgg
     6181 acgtgtaaaa agatatgcat ccagcttcta tatcgcttta actttaccgt tttgggcatc
     6241 gggaacgtat gtaacattga tctcctcttg ggaacggtga gtgcaacgaa tgcgatatag
     6301 caccgaccat gtgggcaaat tcgtaataaa ttcggggtga gggggattca agacaagcaa
     6361 ccttgttagt cagctcaaac agcgatttaa cggttgagta acacatcaaa acaccgttcg
     6421 aggtcaagcc tggcgtgttt aacaagttct tgatatcata tataaatgta ataagaagtt
     6481 tggtaatatt caattcgaag tgttcagtct tttacttctc ttgttttata gaagaaaaaa
     6541 catcaagaaa catctttaac atacacaaac acatactatc agaatacagg atccaaaatg
     6601 gcttctttgg ttaagaaaga tatgtgtatc aagatgacta tggaaggtac tgttaacggt
     6661 catcacttta agtgtgttgg tgaaggtgaa ggtaaaccat ttgaaggtac tcaagttgaa
     6721 aagattagaa ttactgaagg tggtccattg ccatttgctt atgatatttt ggctccatgt
     6781 tgtatgtatg gttctaagac tttcattaag cacgtttctg gtatcccaga ctacttcaaa
     6841 gaatcttttc cagaaggttt tacctgggaa agaactcaaa tctttgaaga tggtggttac
     6901 ttgactattc accaagatac ttcattgcaa ggtaacaact ttatcttcaa agttaacgtt
     6961 atcggtgcta attttccagc taatggtcca gttatgcaaa agaaaactgc tggttgggaa
     7021 ccatgtgttg aaatgttgta tccaagagat ggtgttttgt gcggtcaatc tttgatggct
     7081 ttgaaatgta ctgatggtaa tcacttgacc tctcatttga gaactactta cagatccaga
     7141 aaaccatcta acgctgttaa catgccagaa tttcatttcg gtgaccacag aattgagatt
     7201 ttgaaagctg aacaaggtaa gttctacgaa caatacgaat ctgctgttgc tagatattgt
     7261 gaagctgctc catcaaaatt gggtcatcat taaattgaat tgaattgaaa tcgatagatc
     7321 aatttttttc ttttctcttt ccccatcctt tacgctaaaa taatagttta ttttattttt
     7381 tgaatatttt ttatttatat acgtatatat agactattat ttatctttta atgattatta
     7441 agatttttat taaaaaaaaa ttcgctcctc ttttaatgcc tttatgcagt ttttttttcc
     7501 cattcgatat ttctatgttc gggttcagcg tattttaagt ttaataactc gaaaattctg
     7561 cgttcgt
//
