LOCUS       Exported                6338 bp ds-DNA     circular SYN 18-MAR-2020
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pGAL4-DBD-HEY2wt-IDR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6338)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Mar 18, 2020 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6338
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    944..946
                     /label=start_codon
     CDS             947..1384
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     misc_feature    1394..1942
                     /label=HEY2-wt-IDR
     promoter        complement(1964..1982)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1965..1982)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2002..2019)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer.
                     Also called BGH reverse"
     polyA_signal    2008..2232
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2278..2706
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2365..2384)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2575..2596
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2715..2735)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2720..3050
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2901..3036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2963..2982
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3117..3911
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3171..3190)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3781..3800
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4085..4206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4122..4141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4176..4195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4255..4271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4255..4271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4268..4290)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4279..4295
                     /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(4303..4333)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4348..4369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(4486..4503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4657..5245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4737..4756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5416..6276)
                     /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     6039..6058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcatgaagc tactgtcttc
      961 tatcgaacaa gcatgcgata tttgccgact taaaaagctc aagtgctcca aagaaaaacc
     1021 gaagtgcgcc aagtgtctga agaacaactg ggagtgtcgc tactctccca aaaccaaaag
     1081 gtctccgctg actagggcac atctgacaga agtggaatca aggctagaaa gactggaaca
     1141 gctatttcta ctgatttttc ctcgagaaga ccttgacatg attttgaaaa tggattcttt
     1201 acaggatata aaagcattgt taacaggatt atttgtacaa gataatgtga ataaagatgc
     1261 cgtcacagat agattggctt cagtggagac tgatatgcct ctaacattga gacagcatag
     1321 aataagtgcg acatcatcat cggaagagag tagtaacaaa ggtcaaagac agttgactgt
     1381 atcggcgatc gctcatctca gcacttgcgc cacccagcgg gaggcggcgg ccatgacatc
     1441 ctccatggcc caccaccatc atccgctcca cccgcatcac tgggccgccg ccttccacca
     1501 cctgcccgca gccctgctcc agcccaacgg cctccatgcc tcagagtcaa ccccttgtcg
     1561 cctctccaca acttcagaag tgcctcctgc ccacggctct gctctcctca cggccacgtt
     1621 tgcccatgcg gattcagccc tccgaatgcc atccacgggc agcgtcgccc cctgcgtgcc
     1681 acctctctcc acctctctct tgtccctctc tgccaccgtc cacgccgcag ccgcagcagc
     1741 caccgcggct gcacacagct tccctctgtc cttcgcgggg gcattcccca tgcttccccc
     1801 aaacgcagca gcagcagtgg ccgcggccac agccatcagc ccgcccttgt cagtatcagc
     1861 cacgtccagt cctcagcaga ccagcagtgg aacaaacaat aaaccttacc gaccctgggg
     1921 gacagaagtt ggagcttttt aacgtacgtc tagagggccc tattctatag tgtcacctaa
     1981 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2041 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2101 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2161 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2221 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2281 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2341 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2401 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg
     2461 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2521 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2581 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2641 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2701 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg
     2761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2821 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3001 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3061 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3121 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3181 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3241 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3301 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3361 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3421 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3481 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3541 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3601 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3661 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3721 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3781 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3841 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3901 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3961 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4021 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4081 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4141 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4201 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4261 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4321 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4381 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4441 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4501 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4561 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4621 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4681 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4741 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4801 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     4861 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4921 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4981 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5041 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5101 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5161 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5221 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5281 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5341 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5401 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5461 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5521 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5581 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5641 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5701 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5761 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5821 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5881 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5941 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6001 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6061 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6121 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6181 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6241 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6301 aagcatttat cagggttatt gtctcatgag cggataca
//
