LOCUS       sequence-456934-        4045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For subcloning of human EXOC3 promoter (base pairs -1557 to-1444) or
            for assays using M13 phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4045)
  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 4045)
  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 (-1557 to-1444)
FEATURES             Location/Qualifiers
     source          1..4045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    98..119
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(134..139,140..157,158..164)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    172..188
                     /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     177..199
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     196..212
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     196..212
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(469..488)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(470..488)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(495..512)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(495..511)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(504..526)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      652..1080
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(739..758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     949..970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1424..2218
                     /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(1478..1497)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2088..2107
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2236..2304,2305..3096)
                     /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(2454..2473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3267..3855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3756..3775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4009..4026
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
       61 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      121 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
      181 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt ggtaccgagc
      241 tcggatccac tagtaacggc cgccagtgtg ctggaattcg cccttctcct gcctcagcct
      301 cccgagtagc tgggactcca ggggcccacc accacacctg cctagttttt ttgtattttt
      361 tttagtagag acggggtttc accatgttag ccaggatgga agggcgaatt ctgcagatat
      421 ccatcacact ggcggccgct cgagcatgca tctagagggc ccaattcgcc ctatagtgag
      481 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      541 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      601 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
      661 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
      721 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
      781 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
      841 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
      901 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
      961 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     1021 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     1081 ttaacaaaat tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc
     1141 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa
     1201 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg
     1261 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt
     1321 tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag
     1381 gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     1441 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     1501 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     1561 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     1621 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     1681 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatccca
     1741 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     1801 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     1861 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     1921 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     1981 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     2041 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     2101 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     2161 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat
     2221 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2281 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2341 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2401 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2461 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2521 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2581 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2641 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2701 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2761 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2821 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2881 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2941 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3001 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3061 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3121 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3181 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3241 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3301 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3361 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3421 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3481 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3541 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3601 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3661 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3721 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3781 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3841 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3901 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3961 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4021 agcgaggaag cggaagagcg cccaa
//
