LOCUS       p112A1NE                5725 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  For construction of cDNA expression library..
ACCESSION   .
VERSION     .
KEYWORDS    p112A1NE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5725)
  AUTHORS   Riesmeier JW, Willmitzer L, Frommer WB
  TITLE     Isolation and characterization of a sucrose carrier cDNA from 
            spinach by functional expression in yeast.
  JOURNAL   EMBO J. 1992 Dec;11(13):4705-13.
  PUBMED    1464305
REFERENCE   2  (bases 1 to 5725)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5725
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     113..132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     366..383
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    500..521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        536..566
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    574..590
                     /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)."
     primer_bind     579..601
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     598..614
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     598..614
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        638..1034
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     terminator      1221..1408
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1276..1300)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1420..1437)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1420..1436)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1429..1451)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1873..3219
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3331..4005)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(4006..4107)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4127..4145)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4213..4317
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4318..5178
                     /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     complement(4536..4555)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5349..212
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
       61 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      121 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      181 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      241 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      301 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      361 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
      421 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
      481 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      541 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      601 gaaacagcta tgaccatgat tacgccaagc ttgcatgcaa cttcttttct ttttttttct
      661 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
      721 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
      781 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
      841 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
      901 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      961 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
     1021 gcatacaatc aactccaagc ttcagctgct gcagtctaga gaattcccgg gcggccgcgg
     1081 atccagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct
     1141 tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg
     1201 ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
     1261 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     1321 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
     1381 tcgctcttat tgaccacacc tctaccggca tgccaattca ctggccgtcg ttttacaacg
     1441 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     1501 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     1561 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     1621 acaccgcata tatcggatcg tacttgttac ccatcattga attttgaaca tccgaacctg
     1681 ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat agtctagcgc
     1741 tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc tattgcatag
     1801 gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc acttcaatag
     1861 catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     1921 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     1981 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     2041 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     2101 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     2161 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     2221 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     2281 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     2341 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     2401 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     2461 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     2521 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     2581 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     2641 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     2701 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     2761 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt
     2821 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcggaatag
     2881 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat
     2941 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga
     3001 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta
     3061 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat
     3121 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt
     3181 ctcatccttc aatgctatca tttcctttga tattggatcg atccgatgat aagctgtcaa
     3241 acatgagaat tgatctttta tgcttgcttt tcaaaaggct tgcaggcaag tgcacaaaca
     3301 atacttaaat aaatactact cagtaataac ctatttctta gcatttttga cgaaatttgc
     3361 tattttgtta gagtctttta caccatttgt ctccacacct ccgcttacat caacaccaat
     3421 aacgccattt aatctaagcg catcaccaac attttctggc gtcagtccac cagctaacat
     3481 aaaatgtaag ctctcggggc tctcttgcct tccaacccag tcagaaatcg agttccaatc
     3541 caaaagttca cctgtcccac ctgcttctga atcaaacaag ggaataaacg aatgaggttt
     3601 ctgtgaagct gcactgagta gtatgttgca gtcttttgga aatacgagtc ttttaataac
     3661 tggcaaaccg aggaactctt ggtattcttg ccacgactca tctccatgca gttggacgat
     3721 atcaatgccg taatcattga ccagagccaa aacatcctcc ttaggttgat tacgaaacac
     3781 gccaaccaag tatttcggag tgcctgaact atttttatat gcttttacaa gacttgaaat
     3841 tttccttgca ataaccgggt caattgttct ctttctattg ggcacacata taatacccag
     3901 caagtcagca tcggaatcta gtgcacattc tgcggcctct gtgctctgca agccgcaaac
     3961 tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc aagctgcctt
     4021 tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc cctcttggcc
     4081 ctctcctttt cttttttcga ccgaattaat tcttgaagac gaaagggcct cgtgatacgc
     4141 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4321 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4381 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4441 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4501 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4561 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4621 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4681 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4741 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4801 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4861 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4921 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4981 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5041 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5101 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5161 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5221 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5281 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5341 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5401 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5461 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5521 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5581 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5641 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5701 cgaacgacct acaccgaact gagat
//