LOCUS       sequence-370820-        7186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A parental plasmid for in vivo gene amplification (HamAmp) at SEC23 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7186)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 7186)
  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: pILGFP4BG6
FEATURES             Location/Qualifiers
     source          1..7186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..23
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7..23
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             980..2008
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             980..2008
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2014..2211
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(2958..2991)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3202..3615
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3638..4348
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(4593..4609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4593..4609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4606..4628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4617..4633
                     /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(4641..4647,4648..4665,4666..4671))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4686..4707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4824..4841)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4995..5583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5075..5094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5754..6545,6546..6614))
                     /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     6377..6396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6615..6719)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6787..6805
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6843..6865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6965..6984
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7178..7186,1..14)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gacgttgtaa aacgacggcc agtgaattca gtttaaactc ttctgcttcg ttcagctgca
       61 ggttaaagta gttgtccgtg gacacgagcg tacctctata ttcggtgctg ttgaacttaa
      121 gcttgacgcc tacacgatgg ttgaccaggc ctttgaggaa cggcttcggg ttcaccggct
      181 gcatcgcgct gatatcactg ctctcgctca ttgtcttgtt gtgttgtgac gtagtgtgtg
      241 gtgtggtata gtacgcggtg ctgtgtgtgc tgtgctgttt ttgcgttgct catagaaccg
      301 ttgtttcttg ctcttaccat ctcatgttca tatatgttgg ggaaaagagg tggaacaaga
      361 aaagttaccc ggacggcaga gcggttgttt tgcctgcttt gccataacac aggatgggat
      421 cacacacccc ccaagccatg ctacaacttg gaagagtgca attcaagttg gttgtcctca
      481 gtagtttgag agattgcgct aattttgttt tatcaccgtt atttactgtc agttgagatt
      541 tgtcgcttgc gacgactagc ccgccgcctc tcgtttgtta caaaggagga aagttctagc
      601 aagaataaaa gaaaaacacg tttgaaaaaa gaaaaaaaga tacgggcccg aataccctcc
      661 ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc
      721 ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa
      781 gcaaaaatta cggctcctcg ctgcggacct gcgagcaggg aaacgctccc ctcacagacg
      841 cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac
      901 tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca
      961 catccgaaca taaacaacca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     1021 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     1081 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     1141 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     1201 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
     1261 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     1321 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     1381 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     1441 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     1501 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     1561 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     1621 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     1681 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     1741 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     1801 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     1861 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     1921 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     1981 cagcactcgt ccgagggcaa aggaataatc agtactgaca ataaaaagat tcttgttttc
     2041 aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt
     2101 agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg
     2161 cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt
     2221 cgatactaac ggcggccgcg agcaacggct ttcttttgtt ttatttgatc gttattaggc
     2281 tttttgtata atataaggcc tgttcagtcc attgcgcgtg caagaatttt cctttggacc
     2341 tgaacgaaaa gagtagagga ttttatataa tatattctcc ttcttttttg tactgcgtgg
     2401 cctttactac gaatatctgt atctaatttc ttttctaact tttttattac cattactact
     2461 actactatta ttattattat tattattatt attattgtgt atgactataa tgctttactg
     2521 cctccaatta tgcttttttt attatttttt ttggctgaaa ctacagtata gtaggtatgt
     2581 atatatctaa aactcaaaaa gaggcatgtc taatgtaata ccatcaaaaa caatgaaatt
     2641 atttaatgga gatagccttt tacctagaat gttaatgaca tcctgcgcta cagcaccccc
     2701 tataatggcc gcgacaggag caaactcgat acctttctgt ttgataaact gttgtatgta
     2761 atcgtccttt accactgttg ctggtacacc cagattttca caccatacgg ccgcatctct
     2821 tttcatttgt atttaaatcc atttcaaatt ttatgtttag ttcgagatcc tcagttttcg
     2881 gcgcatagga accacgtaca taataactaa acataaatct ataataaata aaaaacaacg
     2941 atgggagctc gagcctaata acttcgtata gcatacatta tacgaagtta tattaagggt
     3001 tgctcgacca gaaagcgacc acccaacttt ggctgataat agcgtataaa caatgcatac
     3061 tttgtacgtt caaaatacaa tgcagtagat atatttatgc atattacata taatacatat
     3121 cacataggaa gcaacaggcg cgttggactt ttaattttcg aggaccgcga atccttacat
     3181 cacacccaat cccccacaag tgatccccca cacaccatag cttcaaaatg tttctactcc
     3241 ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc
     3301 aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac
     3361 taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca
     3421 ataaaaattt ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt
     3481 tcgatgacct cccattgata tttaagttaa taaacggtct tcaatttctc aagtttcagt
     3541 ttcatttttc ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata
     3601 gcaatctaat ctaagtttta attacaaaat gggatcctct aaaggtgaag aattattcac
     3661 tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt
     3721 ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat ttatttgtac
     3781 tactggtaaa ttgccagttc catggccaac cttagtcact actttcggtt atggtgttca
     3841 atgttttgcg agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc
     3901 agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag
     3961 agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga
     4021 ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata actctcacaa
     4081 tgtttacatc atggctgaca aacaaaagaa tggtatcaaa gttaacttca aaattagaca
     4141 caacattgaa gatggttctg ttcaattagc tgaccattat caacaaaata ctccaattgg
     4201 tgatggtcca gtcttgttac cagacaacca ttacttatcc actcaatctg ccttatccaa
     4261 agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg ctgctggtat
     4321 tacccatggt atggatgaat tgtacaaaag atcttaaact agtaggacta tttgcaaagg
     4381 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
     4441 gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact
     4501 cacaaattag agcttcaatt taattatatc agttattacc cgggaatctc ggtcgtaatg
     4561 atttaaatgc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4621 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4681 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4741 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4801 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4861 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4921 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4981 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5041 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5101 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5161 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5221 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5281 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5341 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5401 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5461 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5521 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5581 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5641 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5701 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5761 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5821 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5881 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5941 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6001 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6061 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6121 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6181 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6241 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6301 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6361 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6421 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6481 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6541 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6601 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6661 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6721 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6781 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     6841 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     6901 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     6961 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     7021 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     7081 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     7141 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcac
//
