LOCUS       sequence-456865-        5032 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For subcloning of human EXOC3 promoter (3' half) or for assays using
            M13 phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5032)
  TITLE     Borchert Lab TOPO plasmids
REFERENCE   2  (bases 1 to 5032)
  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-EXOC3 3' HALF
FEATURES             Location/Qualifiers
     source          1..5032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    109..130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(145..150,151..168,169..175)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    183..199
                     /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     188..210
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     207..223
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     207..223
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1467..1486)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1468..1486)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1493..1510)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1493..1509)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1502..1524)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1650..2078
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1737..1756)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1947..1968
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2422..3216
                     /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(2476..2495)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3086..3105
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3234..3302,3303..4094)
                     /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(3452..3471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4265..4853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4754..4773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5007..5024
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
       61 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      121 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      181 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
      241 tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt gctggaattc gcccttttta
      301 gatctcagga cctgccaccg actcaccctc tacggctacg ccgccgcccc ctccgccttc
      361 gctgcctccg cctccggggt cccggccgga agtggacagc gcgcgggcac ttccgcttcc
      421 ggcaacggcg ccggaagcac gggtccgggc cctcccgcgg ggcgggtact cgtctcccgg
      481 ccgagcgact tcagctcggg gacctgcccc atggccgctc cccagcagga cagggtaacc
      541 gccgaggtgg ggggcttctt gtccgtccca gcccccccaa ctcaggccct ctcgctgccc
      601 ctggcgacca ccgcgtgcgc ccttccccag ggcgggcgca gtcctggacg cctccgccga
      661 gcgtgactgc gtgcattttc tctcctttcc cttcccaacc tgcggagaga agacagcccc
      721 agagccgctg tctccatcct ctgctgatgc ctgctgtctt tatgttggcc tcgtcgtccg
      781 cactgcagtg tggcaggggc gtccctcgtt tcccgcggac tgaggtgggc gccgggcatt
      841 cagtaaacga agaaaccaaa gcggagaagg ttgggaatca aacgtctgtc atacctgcca
      901 ccagcagaca agcggctttg ggtacctcct ggacccaaag acgaactcag cccctccagg
      961 agcgcagtca ttggcaccct cgtgggaaca atgccagtgg gatgggtggc caccggatgt
     1021 ttccggggcc cctcagaggc ccagcagcgc aggtcttgga gaatgaatgt gggtcactgg
     1081 gccgcgctgc agagggccgg tcctgagtcg cttcctggag gagaggccca tggccagcac
     1141 ttggcttcgc ctggaggaag aagcacgtgc taaaccacga gcgttcaggg tttgtccgtt
     1201 tgataaaaga tgctggcttt gacacaccaa ctgacaaagt ctttcatagg ttggagctgc
     1261 taggttgtgc ccctgcccct cctgaaaatt agccacacga aaggaagggt ctggaaggtg
     1321 gcagggaagc cagcagagac cagggaagtg gcagcagcat ggggcagggg cacacaggaa
     1381 ggagatggag cttgcggaag ggcgaattct gcagatatcc atcacactgg cggccgctcg
     1441 agcatgcatc tagagggccc aattcgccct atagtgagtc gtattacaat tcactggccg
     1501 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     1561 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1621 aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc
     1681 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1741 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1801 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1861 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1921 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1981 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2041 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaattc agggcgcaag
     2101 ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg
     2161 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag
     2221 gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc
     2281 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     2341 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag
     2401 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     2461 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2521 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2581 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     2641 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2701 ttgggcgaag tgccggggca ggatctcctg tcatcccacc ttgctcctgc cgagaaagta
     2761 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2821 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     2881 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     2941 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3001 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3061 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3121 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3181 atcgccttct atcgccttct tgacgagttc ttctgaattg aaaaaggaag agtatgagta
     3241 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3301 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3361 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3421 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3481 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3541 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3601 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3661 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3721 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3781 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3841 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3901 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3961 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4021 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4081 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4141 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4201 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4261 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4321 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4381 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4441 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4501 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4561 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4621 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4681 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4741 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4801 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4861 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4921 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4981 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg ga
//
