LOCUS       sequence-452309-        4080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For subcloning of human EXOC3 promoter (base pairs -1592 to -1444) 
            or for assays using M13 phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4080)
  AUTHORS   DeMeis JD, Roberts JT, Delcher HA, Godang NL, Coley AB, Brown CL, 
            Shaw MH, Naaz S, Dahal A, Alqudah SY, Nguyen KN, Nguyen AD, Paudel 
            SS, Shell JE, Patil SS, Dang H, O'Neal WK, Knowles MR, Houserova D, 
            Gillespie MN, Borchert GM
  TITLE     Long G4-rich enhancers target promoters via a G4 DNA-based 
            mechanism.
  JOURNAL   Nucleic Acids Res. 2025 Jan 11;53(2):gkae1180. doi: 
            10.1093/nar/gkae1180.
  PUBMED    39658038
REFERENCE   2  (bases 1 to 4080)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2025-01-11"; volume: "53(2):gkae1180. doi"; 
            pages: " 10.1093/nar/gkae1180"
COMMENT     Sequence Label: TOPO-EXOC3 (-1592 to -1444)
FEATURES             Location/Qualifiers
     source          1..4080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             217..684
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLVPSSDPLVTAASVLEFALSLQAPPPGFTPFSCLSLPS
                     SWDSRGPPPHLPSFFVFFLVETGFHHVSQDGRANSADIHHTGGRSSMHLEGPIRPIVSR
                     ITIHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     complement(513..532)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(514..532)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(539..556)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(539..555)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(548..570)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      696..1124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(783..802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     993..1014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1468..2262
                     /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(1522..1541)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2132..2151
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2280..2348,2349..3140)
                     /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(2498..2517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3311..3899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3800..3819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4053..4070
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgc cctttcactg
      301 caagctccgc ctcccgggtt cacgccattc tcctgcctca gcctcccgag tagctgggac
      361 tccaggggcc caccaccaca cctgcctagt ttttttgtat tttttttagt agagacgggg
      421 tttcaccatg ttagccagga tggaagggcg aattctgcag atatccatca cactggcggc
      481 cgctcgagca tgcatctaga gggcccaatt cgccctatag tgagtcgtat tacaattcac
      541 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
      601 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
      661 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
      721 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      781 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
      841 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      901 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      961 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1021 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1081 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaattcaggg
     1141 cgcaagggct gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa acggtgctga
     1201 ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga
     1261 aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca
     1321 gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa
     1381 gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc aagatctgat
     1441 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     1501 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     1561 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     1621 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     1681 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     1741 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat cccaccttgc tcctgccgag
     1801 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     1861 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     1921 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     1981 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     2041 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     2101 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     2161 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     2221 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaattgaaaa aggaagagta
     2281 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2341 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2401 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2461 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2521 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2581 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2641 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2701 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2761 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2821 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2881 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2941 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3001 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3061 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3121 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3181 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3241 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3301 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3361 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3421 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3481 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3541 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3601 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3661 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3961 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4021 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
