LOCUS       Exported                7075 bp ds-DNA     circular SYN 17-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7075)
  AUTHORS   Dept. Meissner
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 31, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7075
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             710..1150
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     1116..1168
                     /label=Fragment 1.FOR
     misc_feature    1157..1516
                     /label=ROI
     CDS             1277..1279
                     /codon_start=1
                     /label=Feature 2
                     /translation="F"
     CDS             1280..1282
                     /codon_start=1
                     /label=Feature 1
                     /translation="Y"
     CDS             1352..1354
                     /codon_start=1
                     /label=Feature 10
                     /translation="F"
     CDS             1358..1360
                     /codon_start=1
                     /label=Feature 11
                     /translation="F"
     CDS             1397..1516
                     /codon_start=1
                     /label=JAD
                     /translation="PAAPEPLGGICEHETSIDISAYIDPAAFNDEFLADLFQHS"
     CDS             1448..1492
                     /codon_start=1
                     /label=EAD
                     /translation="DISAYIDPAAFNDEF"
     CDS             1460..1462
                     /codon_start=1
                     /label=Feature 3
                     /translation="Y"
     CDS             1478..1480
                     /codon_start=1
                     /label=Feature 12
                     /translation="F"
     primer_bind     complement(1490..1536)
                     /label=Fragment 1.REV
     CDS             1490..1492
                     /codon_start=1
                     /label=Feature 13
                     /translation="F"
     CDS             1493..1504
                     /codon_start=1
                     /label=Feature 100
                     /translation="LADL"
     primer_bind     1497..1538
                     /label=Fragment 2.FOR
     CDS             1505..1507
                     /codon_start=1
                     /label=Feature 14
                     /translation="F"
     misc_feature    1517..2377
                     /label=FUS_IDR
     primer_bind     complement(2349..2417)
                     /label=Fragment.REVa
     promoter        complement(2394..2412)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2395..2412)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2432..2449)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2438..2662
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2708..3136
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2795..2814)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3005..3026
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3145..3165)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3150..3479
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3330..3465
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3392..3411
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3546..4340
                     /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(3600..3619)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4210..4229
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4514..4635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4551..4570)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4605..4624
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4684..4700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4684..4700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4697..4719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4708..4724
                     /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(4732..4762)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4777..4798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4915..4932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5086..5674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5166..5185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5845..6705)
                     /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     6468..6487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6706..6810)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6885..6904)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattca tgaagctact
      721 gtcttctatc gaacaagcat gcgatatttg ccgacttaaa aagctcaagt gctccaaaga
      781 aaaaccgaag tgcgccaagt gtctgaagaa caactgggag tgtcgctact ctcccaaaac
      841 caaaaggtct ccgctgacta gggcacatct gacagaagtg gaatcaaggc tagaaagact
      901 ggaacagcta tttctactga tttttcctcg agaagacctt gacatgattt tgaaaatgga
      961 ttctttacag gatataaaag cattgttaac aggattattt gtacaagata atgtgaataa
     1021 agatgccgtc acagatagat tggcttcagt ggagactgat atgcctctaa cattgagaca
     1081 gcatagaata agtgcgacat catcatcgga agagagtagt aacaaaggtc aaagacagtt
     1141 gactgtatcg gcgataatgc gcgggcgcgg gcgagcaggg tctccgggtg ggcggcggag
     1201 gagacctgca caagcaggcg gaaggcgagg atccccgtgt cgcgaaaatt ctaattctcc
     1261 aatggaaagc gctgatttct atgaagcaga acccaggcca ccaatgtctt cccatctcca
     1321 aagtccccct catgcaccaa gttccgcagc atttgggttc cctagaggag caggaccagc
     1381 ccaaccccct gctcccccgg ctgctcccga acctctcggt ggaatttgtg aacatgaaac
     1441 cagcatagat atttctgcat atatagatcc agctgctttt aatgatgaat ttcttgcgga
     1501 tctctttcaa cattctgcgt ccaatgacta cactcagcag gccacacagt cttacggtgc
     1561 atatccaaca caaccaggac aagggtacag ccaacaaagt tctcaaccat atggccaaca
     1621 atcctatagc ggctactcac aatcaacaga taccagtggg tacgggcaaa gtagttactc
     1681 aagctacgga caatcccaaa ataccggtta cggcacacaa tccacacctc aagggtacgg
     1741 ttccacaggt ggttacggaa gctcccaatc cagtcagagc agctatggcc aacaaagcag
     1801 ctaccccgga tacgggcaac aaccggcccc ttcttccaca agcggctcct atggatcaag
     1861 ctcccaatct agttcttacg gccaaccgca atccggctct tatagtcaac aacccagtta
     1921 cggcggtcaa caacagagtt acggccaaca acagtcctac aacccgccac aagggtacgg
     1981 ccaacaaaat caatataata gtagttctgg aggaggcggg gggggcggcg gtggtggcaa
     2041 ttacgggcag gaccagagct ctatgagctc cggtggcggt tccggcggtg ggtacgggaa
     2101 ccaagatcaa tccggcggtg gtggttccgg tggatacggc caacaagatc ggggcggtcg
     2161 agggagaggt ggatctggag gaggtggtgg aggagggggg ggcggctata atagatcttc
     2221 tggtggatac gagccacgcg gccgcggcgg cggacgaggc gggcgtggtg gaatgggtgg
     2281 ctccgatagg ggcggtttta acaagttcgg cggacccagg gatcagggca gtcgccacga
     2341 tagcgagcag gataattcag acaacaacac catctgagtc tagagggccc tattctatag
     2401 tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     2461 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2521 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2581 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2641 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2701 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2761 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2821 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2881 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2941 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3001 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3061 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3121 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3181 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3301 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3361 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3421 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3481 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3541 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3601 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3661 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3721 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3781 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3841 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3901 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3961 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4021 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4081 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4141 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4201 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4261 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4321 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4381 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4441 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4501 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4561 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4621 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5881 gatctgtcta tttcgttcat ccatagttgc ctgacttccc gtcgtgtaga taactacgat
     5941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6841 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     6901 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     6961 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7021 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
