LOCUS       sequence-51351-e        5705 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5705)
  TITLE     pFA6-yEGFP1-HIS3MX6
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5705)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pFA6-luc*(-SKL)-HIS3MX6
FEATURES             Location/Qualifiers
     source          1..5705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             61..1710
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKKL"
     primer_bind     complement(127..145)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      1731..1918
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1786..1810)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1993..2336
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2337..3035
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      3039..3236
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3340..3359)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3341..3359)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3446..3463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3617..4205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3697..3716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4376..5167,5168..5236))
                     /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     4999..5018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5237..5341)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5409..5427
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     5587..5606
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5687..5705
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5687..5704
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct agaggatgga
      121 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt
      181 gcttttacag atgcacatat cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc
      241 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta
      301 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt
      361 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgaacatt
      421 tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa
      481 aaaaaattac caataatcca gaaaattatt atcatggatt ctaaaacgga ttaccaggga
      541 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat
      601 tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa ctcctctgga
      661 tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt cagattctcg
      721 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt
      781 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt
      841 cgagtcgtct taatgtatag atttgaagaa gagctgtttt tacgatccct tcaggattac
      901 aaaattcaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa aagcactctg
      961 attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc acctctttcg
     1021 aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg acaaggatat
     1081 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc
     1141 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa
     1201 acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat tatgtccggt
     1261 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct
     1321 ggagacatag cttactggga cgaagacgaa cacttcttca tagttgaccg cttgaagtct
     1381 ttaattaaat acaaaggata ccaggtggcc cccgctgaat tggagtcgat attgttacaa
     1441 caccccaaca tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc cggtgaactt
     1501 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat
     1561 tacgtcgcca gtcaagtaac aaccgccaaa aagttgcgcg gaggagttgt gtttgtggac
     1621 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata
     1681 aaggccaaga agggcggaaa gaagctttaa ggcgcgccac ttctaaataa gcgaatttct
     1741 tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt
     1801 aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg
     1861 tcaggttgct ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca
     1921 gatccgctag ggataacagg gtaatataga tctgtttagc ttgcctcgtc cccgccgggt
     1981 cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc
     2041 tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca
     2101 tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg
     2161 cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg
     2221 cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt
     2281 ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg
     2341 cagaaccagc ccaaaaaaag caaaaacaaa ctgttcagga gcgcaaggcg tttatctccc
     2401 gtatcactaa tgaaactaaa attcaaatcg ctatttcgct gaatggtggt tatattcaaa
     2461 taaaagattc gattcttcct gcaaagaagg atgacgatgt agcttcccaa gctactcagt
     2521 cacaggtcat cgatattcac acaggtgttg gctttttgga tcatatgatc catgcgttgg
     2581 caaaacactc tggttggtct cttattgttg aatgtatcgg tgacctgcac attgacgatc
     2641 accatactac cgaagattgc ggtatcgcat tagggcaagc gttcaaagaa gcaatgggtg
     2701 ctgtccgtgg tgtaaaaaga ttcggtactg ggttcgcacc attggatgag gcgctatcac
     2761 gtgccgtagt cgatttatct aatagaccat ttgctgtaat cgaccttgga ttgaagagag
     2821 agatgattgg tgatttatcc actgaaatga ttccacactt tttggaaagt ttcgcggagg
     2881 cggccagaat tactttgcat gttgattgtc tgagaggttt caacgatcac cacagaagtg
     2941 agagtgcgtt caaggctttg gctgttgcca taagagaagc tatttctagc aatggcacca
     3001 atgacgttcc ctcaaccaaa ggtgttttga tgtgaagtac tgacaataaa aagattcttg
     3061 ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa
     3121 atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa
     3181 gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt
     3241 cgattcgata ctaacgccgc catccagttt aaacgagctc gaattcatcg atgatatcag
     3301 atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa
     3361 tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3421 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3481 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3541 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3601 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3661 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3721 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3781 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     3841 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3901 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3961 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4021 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4081 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4141 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4201 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4261 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4321 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4381 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4441 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4501 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4561 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4621 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4681 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4741 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4801 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4861 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4921 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4981 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5041 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5101 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5161 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5221 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5281 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5341 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5401 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     5461 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     5521 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     5581 actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag
     5641 aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact
     5701 ataga
//
