LOCUS       sequence-374392-       10094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo gene amplification (HamAmp) of AeBlue + HPV16 L1 expression 
            module at RPL25 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10094)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 10094)
  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: pIAeBlueHPV16LR
FEATURES             Location/Qualifiers
     source          1..10094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1974..1992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2030..2052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2152..2171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2365..2387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2379..2396
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2380..2396
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6290..6560
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     CDS             8754..8771
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9874..9890)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9874..9890)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9887..9909)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9898..9914
                     /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(9922..9928,9929..9946,9947..9952))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9967..9988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2221 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2281 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2341 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     2401 tcagtttaaa catgtactaa tcagtctaac accccggagg aggcttatca ttacatattc
     2461 ttgtctcggc gttctttaag tttcttatct tgtatgcccg atatagcaac cttgttggta
     2521 ccaatctaac ggtttccgta ctttgcaatg aagagatgag gaggcatggg tcacttattt
     2581 aatatgtacg ggtgtttaca tggagttgct ttcttttttt gtctcagcag tcattgtgcg
     2641 ccaaaaaaag agaaaaccgt gagccgaagt ccacgctctg gagttaggct ctcccattac
     2701 ggagagaagc atttcctcag cctgggagcc ccgttggaac agtcaggcta aactgggcct
     2761 tcctacccac tgcttgctgt ttctcactgg acgcacaagg ggattttctt tctaccttcg
     2821 gcttgcctca ctgcgttggg gcttcccaat gcaacttcgt tcgtatgcat acaatctttt
     2881 agatattatc ttttaaaatt attttaaaac aattttaaat gtatctcata tgcttttctt
     2941 ctgctgttga aaaggctaaa caaagaagat caataagata aagcggccgc aaaatgtcta
     3001 agaatatcgt tgtcctaccg ggtgatcacg tcggtaaaga agttactgac gaagctatta
     3061 aggtcttgaa tgccattgct gaagtccgtc cagaaattaa gttcaatttc caacatcact
     3121 tgatcggggg tgctgccatc gatgccactg gcactccttt accagatgaa gctctagaag
     3181 cctctaagaa agccgatgct gtcttactag gtgctgttgg tggtccaaaa tggggtacgg
     3241 gcgcagttag accagaacaa ggtctattga agatcagaaa ggaattgggt ctatacgcca
     3301 acttgagacc atgtaacttt gcttctgatt ctttactaga tctttctcct ttgaagcctg
     3361 aatatgcaaa gggtaccgat ttcgtcgtcg ttagagaatt ggttggtggt atctactttg
     3421 gtgaaagaaa agaagatgaa ggtgacggag ttgcttggga ctctgagaaa tacagtgttc
     3481 ctgaagttca aagaattaca agaatggctg ctttcttggc attgcaacaa aacccaccat
     3541 taccaatctg gtctcttgac aaggctaacg tgcttgcctc ttccagattg tggagaaaga
     3601 ctgttgaaga aaccatcaag actgagttcc cacaattaac tgttcagcac caattgatcg
     3661 actctgctgc tatgattttg gttaaatcac caactaagct aaacggtgtt gttattacca
     3721 acaacatgtt tggtgatatt atctccgatg aagcctctgt tattccaggt tctttgggtt
     3781 tattaccttc tgcatctcta gcttccctac ctgacactaa caaggcattc ggtttgtacg
     3841 aaccatgtca tggttctgcc ccagatttac cagcaaacaa ggttaaccca attgctacca
     3901 tcttatctgc agctatgatg ttgaagttat ccttggattt ggttgaagaa ggtagggctc
     3961 ttgaagaagc tgttagaaat gtcttggatg caggtgtcag aaccggtgac cttggtggtt
     4021 ctaactctac cactgaggtt ggcgatgcta tcgccaaggc tgtcaaggaa atcttggctt
     4081 aaagagtctt ttgtaacgac cccgtctcca ccaacttggt atgcttgaaa tctcaaggcc
     4141 attacacatt cagttatgtg aacgaaaggt ctttatttaa cgtagcataa actaaataat
     4201 acaggttccg gttagcctgc gggcccaaag ctagctctaa ttggtttaat taataaattt
     4261 aatattattt ttaaattttt ctttaaatat acaataaatc tttcataaca tgttaaattc
     4321 atgattaagc gtaaataaag tgtagtggca gagtgcacgg ggtttcctgt gccttacaaa
     4381 gtaggtacca atttgcgtat tgcagcgagg gttccggtta ctatttataa ttacgtgtta
     4441 gtgtactgtg attttattga ggctataaca agaaaaggat ctgtgaaggt tttgagagaa
     4501 tgaggcgaaa acacagggtt attttcatca gcaacgccgc tgtttcagcg gcaccctgcc
     4561 cttttctttt cgcaagatga gagtaaagag ttgtacatca ggtaagaatg ttattattta
     4621 aattcgaagt gataaattct tttcatgatg aatcactcgc ttatatgggg tagaatatat
     4681 atatatgtgt gtgtgtgtgt gtgtttgtgt atgtagggat ggtgcgcgtt tgttgtgtga
     4741 catttgctac tcattctttt ccttttccta cgactggctt aacgggaata ttatcaattt
     4801 gctgcattct tatgcttcgg tccgatgctc aagcttaact tcttcgtgag gaaggaaagt
     4861 gttgtctcct acttttttca aattttcgaa ttgtatttat atttatttag tacttcttga
     4921 gtttacatat ccttcgtaaa aatgcaactt ttgtcgaaaa acacttccaa aaaaaaataa
     4981 taatgaattt atgaagcata ctaacgagcg agcacatcgc tgaggtatca ttacttcatg
     5041 agataaatta actcgagcat atggcgtatc caagccgaaa cggcgctcgc ctcatcccca
     5101 cgggaataag gcagccgaca aaagaaaaac gaccgaaaag gaaccagaaa gaaaaaagag
     5161 ggtgggcgcg ccgcggacgt gtaaaaagat atgcatccag cttctatatc gctttaactt
     5221 taccgttttg ggcatcggga acgtatgtaa cattgatctc ctcttgggaa cggtgagtgc
     5281 aacgaatgcg atatagcacc gaccatgtgg gcaaattcgt aataaattcg gggtgagggg
     5341 gattcaagac aagcaacctt gttagtcagc tcaaacagcg atttaacggt tgagtaacac
     5401 atcaaaacac cgttcgaggt caagcctggc gtgtttaaca agttcttgat atcatatata
     5461 aatgtaataa gaagtttggt aatattcaat tcgaagtgtt cagtctttta cttctcttgt
     5521 tttatagaag aaaaaacatc aagaaacatc tttaacatac acaaacacat actatcagaa
     5581 tacaggatcc aaaatggctt ctttggttaa gaaagatatg tgtatcaaga tgactatgga
     5641 aggtactgtt aacggtcatc actttaagtg tgttggtgaa ggtgaaggta aaccatttga
     5701 aggtactcaa gttgaaaaga ttagaattac tgaaggtggt ccattgccat ttgcttatga
     5761 tattttggct ccatgttgta tgtatggttc taagactttc attaagcacg tttctggtat
     5821 cccagactac ttcaaagaat cttttccaga aggttttacc tgggaaagaa ctcaaatctt
     5881 tgaagatggt ggttacttga ctattcacca agatacttca ttgcaaggta acaactttat
     5941 cttcaaagtt aacgttatcg gtgctaattt tccagctaat ggtccagtta tgcaaaagaa
     6001 aactgctggt tgggaaccat gtgttgaaat gttgtatcca agagatggtg ttttgtgcgg
     6061 tcaatctttg atggctttga aatgtactga tggtaatcac ttgacctctc atttgagaac
     6121 tacttacaga tccagaaaac catctaacgc tgttaacatg ccagaatttc atttcggtga
     6181 ccacagaatt gagattttga aagctgaaca aggtaagttc tacgaacaat acgaatctgc
     6241 tgttgctaga tattgtgaag ctgctccatc aaaattgggt catcattaaa ttgaattgaa
     6301 ttgaaatcga tagatcaatt tttttctttt ctctttcccc atcctttacg ctaaaataat
     6361 agtttatttt attttttgaa tattttttat ttatatacgt atatatagac tattatttat
     6421 cttttaatga ttattaagat ttttattaaa aaaaaattcg ctcctctttt aatgccttta
     6481 tgcagttttt ttttcccatt cgatatttct atgttcgggt tcagcgtatt ttaagtttaa
     6541 taactcgaaa attctgcgtt cgttaaagct ttcgagaagg atattatttc ccgggccaca
     6601 gagaacagga gattacaaga tccagatcac caaaatgccg gaacacaggc tgaaaaactc
     6661 caagcagccg cacacccgat ttttagccag cagattcata cctaactcca atgaattgca
     6721 gcatatatcc tgcaataaca cattaaggat cgggccttgg acggccgaat atatgattat
     6781 agtccttttg tcgccaaaaa gtcgcctcga ggtcacgtgc atccgaaatc accccctgag
     6841 cacttcccgc ccagaaaaca gggaaggggc ggcaccagca ggcaccggca ttcactcgtc
     6901 cgtgcggcgt ggtctgtgcc gttcaggggc ctgcgtgcct gtgatgatat cgctgcaggg
     6961 caccgagtcg gagaggacca ctccgagccg aggttagtta agccattctc tacgctaaaa
     7021 aataaataat gtggattaag ctcctactgg aaggttaagt gtttgtacac tttcaaacat
     7081 gcgcaacgca ccaggcaact attgttctga ggagggatgt gatgcggagt aaaatgtgga
     7141 ggaattacgg ggcttgcaga catatataaa ctctagccat tcggctgaat ataagctaga
     7201 aaccacttct tctctttcca tctacatctc cctcatctat caataagaac tataatatca
     7261 acaatactat aatataatat atacacacat ttacagtcga caaaatgtct ttgtggttgc
     7321 catctgaagc tactgtttac ttgccaccag ttccagtttc taaggttgtt tctactgatg
     7381 aatacgtcgc cagaactaac atctattatc atgctggtac ttccaggttg ttggctgttg
     7441 gtcatccata ttttccaatc aagaagccca acaacaacaa gattttagtt ccaaaagttt
     7501 ccggtttaca gtacagagtt ttcagaatcc atttgccaga tccaaacaag tttggtttcc
     7561 cagatacttc attctacaac ccagatacac aaagattggt ttgggcttgt gttggtgttg
     7621 aagttggtag aggtcaacca ttaggtgttg gtatttctgg tcatcctttg ttgaacaagt
     7681 tggacgatac tgaaaatgct tctgcttatg ctgctaatgc tggtgttgat aacagagaat
     7741 gcatctctat ggattacaag caaacccagt tgtgtttgat tggttgtaaa ccacctattg
     7801 gtgaacattg gggtaaaggt tctccatgta ctaatgttgc tgttaatcca ggtgactgtc
     7861 caccattgga attgattaac accgttattc aagatggcga tatggttgat actggttttg
     7921 gtgctatgga tttcactaca ttgcaagcca acaaatccga agttccattg gatatctgta
     7981 cctctatctg taagtaccca gactacatca agatggttag tgaaccttat ggtgactcct
     8041 tgttcttcta cttgagaaga gaacaaatgt tcgtcaggca tttgtttaat agagctggtg
     8101 ctgttggtga aaatgttcca gatgacttgt acatcaaagg ttctggttct actgctaact
     8161 tggcttcttc taattacttc ccaactccat ctggttcaat ggttacttct gatgcccaaa
     8221 ttttcaacaa gccatattgg ttgcaaagag cccaaggtca taacaatggt atttgttggg
     8281 gtaaccagtt gttcgttact gttgttgata ccaccagatc cactaacatg tctttatgtg
     8341 ctgctatctc tacttccgaa actacttaca agaacacgaa cttcaaagaa tacttgaggc
     8401 acggtgaaga atacgacttg caattcattt ttcagctgtg caagattacc ttgaccgctg
     8461 atgttatgac ttacatccat tctatgaact ccaccatctt ggaagattgg aattttggtc
     8521 tacaaccacc accaggtggt actttggaag atacctatag attcgttacc tctcaagcta
     8581 tcgcttgtca aaaacatact ccaccagctc caaaagaaga tccattgaag aaatacactt
     8641 tctgggaagt caacctgaaa gaaaagttct ctgctgattt ggatcaattc ccattgggta
     8701 gaaagttctt gttgcaagct ggtttgaaag ctaagccaaa gttcactttg ggtcaccacc
     8761 atcaccatca ttaatagtga cctgcagggc ggattgagag caaatcgtta agttcaggtc
     8821 aagtaaaaat tgatttcgaa aactaatttc tcttatacaa tcctttgatt ggaccgtcat
     8881 cctttcgaat ataagatttt gttaagaata ttttagacag agatctactt tatatttaat
     8941 atctagatat tacataattt cctctctaat aaaatatcat taataaaata aaaatgaagc
     9001 gatttgattt tgtgttgtca acttagtttg ccgctatgcc tcttgggtaa tgctattatt
     9061 gaatcgaagg gctttattat attacccttt agcttattct gaggtttctg tggcgtgccg
     9121 gcaatctcgg tcgtaatgat ttgcatgccc gccatctcta ctcactccat aatattacat
     9181 atagatatag gacaagcccg cattttcata ctgaaaggta aacttctatt attatagtgg
     9241 tatccaacgt tcaccgcttc cagcatagca gaaattacgt gtttttgcat atgttatgct
     9301 gatcattgta tgcttactac catttttctt tgcttcgcct tgccttcttt gacgtttttt
     9361 tgaagcaaaa aaaaagtcaa gacagatgtg cttacaaaac catgtaaggc tcattttcaa
     9421 agaagctact aatagaaaga gaacaaagcg tttacgagtc tggaaaatca atggctccat
     9481 ctggtatgtg aactgcaata ttaatagcac gagaaaattg agaggaagat agatgggaac
     9541 tagtagagtt gatattgatg agatacgaaa accacacgta aataaactat ccgacgacaa
     9601 gaatagtgca ttaaggactt gattaagata tggtgagcaa cgtaattatc gggctcaaca
     9661 gtttattagc aatcgttttg atagaagcgt tgatgctgtg gaagttgttc tttttactac
     9721 caacagccat taacaaatcc attagagtgt tcgttcgttt ttctgctcag attcaagaaa
     9781 attattccat cctcattata cttttttctt ctatttcgtg ctccacgtcg aggtatcaag
     9841 gaacatagtt taaacaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     9901 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     9961 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    10021 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    10081 gcgtattggg cgct
//
