LOCUS       sequence-485897-        4443 bp    DNA     circular SYN 12-SEP-2025
DEFINITION  For subcloning of human EXOC3 promoter (base pairs -1557 to-1046) or
            for assays using M13 phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4443)
  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 4443)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 12, 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-1046)
FEATURES             Location/Qualifiers
     source          1..4443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(698..714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(698..714)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(711..733)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    722..738
                     /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(join(746..752,753..770,771..776))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    791..812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(929..946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1100..1688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1180..1199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1859..2650,2651..2719))
                     /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     2482..2501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(2737..3531)
                     /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(2848..2867)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3458..3477
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(3875..4303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3985..4006)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4197..4216
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(4429..4443,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4443,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg ggccctctag
       61 atgcatgctc gagcggccgc cagtgtgatg gatatctgca gaattcgccc ttctcctgcc
      121 tcagcctccc cagtagctgg gactccaggg gcccaccacc acacctgcct agtttttttg
      181 tatttttttt agtagagacg gggtttcacc atgttagcca ggacggtctc gatctcctga
      241 cctcatgatc tacctgcctc gcctcccaaa gtgctgggat tacaggcgtg agccacggcg
      301 ccccgccaag aaagacttct aatcaatgtt ttgttaagaa gttactagac agtcctccac
      361 ccaaattcac taccagacac tgatataata gtatgagtct ttagttcata atactcaatg
      421 ttactgtatt tttaagagtc ctaaaagctt tggcaatttt ggcaattttt tcttcattag
      481 tttcaaaatt tctttacagt cataatttct aacacaattt taactttttt actttttttc
      541 attgctcaac ttgttgctag attttcagag attacaagga aaaactgaaa taatagagcc
      601 ggggtcagtg tcaacatcag caggaagggc gaattccagc acactggcgg ccgttactag
      661 tggatccgag ctcggtacca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      721 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      781 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      841 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      901 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
      961 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1021 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1261 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1501 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1981 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2161 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2461 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2701 ggaaatgttg aatactcata ctcttccttt ttcaattcag aagaactcgt caagaaggcg
     2761 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     2821 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     2881 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     2941 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     3001 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     3061 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     3121 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     3181 agccatgatg gatactttct cggcaggagc aaggtgggat gacaggagat cctgccccgg
     3241 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     3301 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     3361 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     3421 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     3481 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     3541 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     3601 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
     3661 caattccggt tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc
     3721 actgcaagct acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta
     3781 gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt
     3841 cctttagcag cccttgcgcc ctgaattttg ttaaaattcg cgttaaattt ttgttaaatc
     3901 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3961 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4021 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4081 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4141 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4201 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4261 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     4321 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     4381 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     4441 gtt
//
