LOCUS       sequence-455930-        6000 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For subcloning of human ZNF84 promoter or for assays using M13 
            phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6000)
  TITLE     Borchert Lab TOPO plasmids
REFERENCE   2  (bases 1 to 6000)
  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"
COMMENT     Sequence Label: TOPO-ZNF84
FEATURES             Location/Qualifiers
     source          1..6000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    87..108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(123..128,129..146,147..153)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    161..177
                     /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     166..188
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     185..201
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     185..201
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2413..2432)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2414..2432)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2439..2456)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2439..2455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2448..2470)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2596..3024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2683..2702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2893..2914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3368..4162
                     /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(3422..3441)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4032..4051
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4180..4248,4249..5040)
                     /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(4398..4417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5211..5799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5700..5719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5953..5970
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
       61 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      121 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      181 cacacaggaa acagctatga ccatgattac gccaagcttg gtaccgagct cggatccact
      241 agtaacggcc gccagtgtgc tggaattcgc ccttcttacc tataattctc cctacgactc
      301 cagttatgga tcaactttac ttttttctgt taattcttac cttctttcca taaatatatt
      361 taaatttttt ctggtttcct caactgaagt caaaccaatt gagtattggt gggccgggcg
      421 cggtggctca cgcctgtaat cctagcactt tgggaggcca aggcgggcgg attgcctgag
      481 ctcaggagtt cgcgaccagc ctgggcaaca cggtgactcc ccgtctctac taaaatacaa
      541 aaaaattagc cgggcgtggc ggcgggcgcc tgtagtccca gctactcggg aggctgaggc
      601 agaagaatgg cgtcaacccg ggaggcggag cttgcagtga gccgagatca cgccactgca
      661 ctccagcctg ggtgacagag cgagacgccg tctcaaaaaa atatatatat atattggtgg
      721 attcggtttc cctttatgtt tctcagaaaa ccgtgtgagc ccgtttggag tcactgcgtc
      781 tccaagcgac aggcacttcc ggacgcgggg cactgtggga agttgagttc gcggggctcc
      841 ttccttcgcg catgcgcaca tgcactcttg gtaatgcttg caggtggccc gttcttcctc
      901 tttgtccttc gctgccacgt gagaggggag tgtacgcgtg tgcgcgcgtg cgagtgtgcg
      961 cgtgtgcagt ccccgactcg tcccggcccg ttcggagccg cccgagagcg cacaaaggcc
     1021 gacggtttgg gcgcgtgcgc gggacacggg ggacgggcgg tgtcacgtga tcgcgaggcc
     1081 gcgggcgcgc gggcgcgact cggcggcggc gtcttttgtg agtccagtga gtttattgcg
     1141 ggcccggcgc tgggggctcc gggaatggcg ggggaggcgc gcgggagtgg ccgcggggag
     1201 cttcggcttc aaggccctgg gcgtccgcgc gcggatgcgg gtctgggacg ggttggggct
     1261 gggccgggtc gtgaagatgc cgtgggccgc ggagctgcgc ggattctggg ctgtggggcg
     1321 ggcacggcct ttccggaggc gccgctcctg ggcgtccttt caggcgggcg tggaggcgcc
     1381 agcctcgggc gcgtttagcc tcgacctcgg gccagcccgc ttgccgcgcc gtggccctaa
     1441 ggagcggctg ccctgctgga ccgcatccac cagcagcctg gctctgccgc tatcgaagga
     1501 ggcttttaaa gcgctggggt tacaggtgtg agccaccatt cctggccgtg tattttttaa
     1561 tggttatatt tattgactgt tttaaatttt ttaagtgact tttgcttttg gtatggtgta
     1621 gcgaaatctc ccctcaaatt atacatatat tcattcacct gtggtttttt ttgtagacat
     1681 tcttagggtt tccattattt taaaatttaa cccactgaaa gtagttgtat gaaatagcta
     1741 cgtttatttt ctttcaaata ttcttgtatt cattaaataa tttaatttta ctgatggctt
     1801 tgaagtgcta catttattgc atgctgactc ttgcatattt agacagttct gctttttttc
     1861 atctgtataa cttctctgga gccagtgtca aatgttataa tgattataat ttgtatgtat
     1921 tacagtgggc ctgttcacac tcattatttt gtatttccaa aagcatccag gcccggtgtg
     1981 gtagggcatg cctgtagccc cagctactct gtaggttgag gtaggaggat cacttgagcc
     2041 caggagttca gggctgcagt gggctatggt tatgccattg cacttcagcc tggagaacac
     2101 agtaagaccc tgtcaaaaaa aaaaaaaggt gtccgaattg ttcttgaacc ttttttctta
     2161 cacataaact ttagaatttt gagattctga tcggagtcga atgagttata taaatcattt
     2221 gaggtcaaat tatcttcaca atctgaagtc ttcctatccc agaataaaat cgaattataa
     2281 atttctttaa ttctattttt atagttctca ttggtgtttt tagtatgctt catatgggtc
     2341 ttgaagggcg aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga
     2401 gggcccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     2461 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     2521 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     2581 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2641 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2701 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2761 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2821 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2881 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2941 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3001 tttaacaaaa atttaacgcg aattttaaca aaattcaggg cgcaagggct gctaaaggaa
     3061 gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta
     3121 ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg
     3181 gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc
     3241 agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt
     3301 gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc
     3361 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3421 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3481 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3541 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3601 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3661 ggggcaggat ctcctgtcat cccaccttgc tcctgccgag aaagtatcca tcatggctga
     3721 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3781 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3841 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3901 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3961 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4021 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4081 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4141 ccttcttgac gagttcttct gaattgaaaa aggaagagta tgagtattca acatttccgt
     4201 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4261 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4321 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4381 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4441 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4501 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4561 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4621 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4681 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4741 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4801 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4861 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4921 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4981 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5041 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5101 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5161 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5221 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5281 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5341 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5401 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5461 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5521 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5581 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5641 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5701 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5761 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5821 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5881 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5941 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
//
