LOCUS       Exported                7036 bp ds-DNA     circular SYN 12-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7036)
  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..7036
                     /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.FOR
     primer_bind     complement(2323..2378)
                     /label=Fragment.REV
     promoter        complement(2355..2373)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2356..2373)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2393..2410)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2399..2623
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2669..3097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2756..2775)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2966..2987
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3106..3126)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3111..3440
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3291..3426
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3353..3372
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3507..4301
                     /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(3561..3580)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4171..4190
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4475..4596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4512..4531)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4566..4585
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4645..4661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4645..4661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4658..4680)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4669..4685
                     /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(4693..4723)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4738..4759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4876..4893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5047..5635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5127..5146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5806..6666)
                     /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     6429..6448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6667..6771)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6846..6865)
                     /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 gcgatcatgc 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 cattctaggc aacaagagaa agcaaaagct gcggtaggac caactggtgg
     1561 tggtggagga ggagatttcg attatcctgg tgccccagca ggtcctggtg gggcagtaat
     1621 gccgggtggg gcacatggac caccccctgg gtatgggtgt gctgcagctg ggtatcttga
     1681 tggacgactt gaaccacttt atgaacgggt gggagctccc gcactcaggc ccctcgtcat
     1741 aaaacaagaa cctcgggaag aagacgaggc taaacaactt gctcttgctg ggctgtttcc
     1801 gtatcaaccc ccacccccac caccaccatc ccatcctcat ccccatccac cacctgctca
     1861 tttggcggca ccacatttgc aatttcaaat tgcacattgt gggcaaacta ctatgcatct
     1921 tcaacctggc catccgacac ccccaccaac cccggttcca agtccacatc ctgcacctgc
     1981 tctgggagca gctggactcc ctggtccggg gtctgccttg aaaggcctcg gtgcagcaca
     2041 tccagatctg agagcttcag gcgggtccgg ggcaggaaaa gcaaagaaaa gtgtcgataa
     2101 gaatagtaat gaatatagag ttcgccgtga aaggaataat atagccgtcc ggaaatctag
     2161 ggataaagct aaacaacgaa atgtcgaaac ccaacaaaag gttctcgaac tcactagcga
     2221 taacgatagg ctcagaaaac gtgtagagca actctctcgg gagttggata ccttgagagg
     2281 catcttccgc cagctgccag agagctcctt ggtcaaggcc atgggcaact gcgcgtgagt
     2341 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact
     2401 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2461 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2521 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2581 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2641 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2701 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2761 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2821 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2881 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2941 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3001 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3061 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3121 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3181 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3241 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3301 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3361 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3421 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3481 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3541 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3601 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3661 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3721 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3781 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3841 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3901 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3961 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4021 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4081 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4141 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4201 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4261 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4321 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4381 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4441 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4501 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4561 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4621 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4681 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4741 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4801 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4861 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4921 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4981 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5041 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5101 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5161 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5221 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5281 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5341 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5401 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5461 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5521 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5581 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5641 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5701 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5761 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5821 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgacttcc
     5881 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5941 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6001 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6061 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6121 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6181 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6241 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6301 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6361 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6421 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6481 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6541 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6601 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6661 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6721 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6781 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     6841 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     6901 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     6961 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     7021 gccagatata cgcgtt
//
