LOCUS       sequence-456933-        4568 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For subcloning of human EXOC3 promoter (base pairs -2455 to -1951) 
            or for assays using M13 phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4568)
  TITLE     Borchert Lab TOPO plasmids
REFERENCE   2  (bases 1 to 4568)
  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 (-2455 to -1951)
FEATURES             Location/Qualifiers
     source          1..4568
                     /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"
     primer_bind     complement(1001..1020)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1002..1020)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1027..1044)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1027..1043)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1036..1058)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1184..1612
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1271..1290)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1481..1502
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1956..2750
                     /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(2010..2029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2620..2639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2768..2836,2837..3628)
                     /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(2986..3005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3799..4387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4288..4307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4541..4558
                     /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 ccttaaaaaa
      301 aaaaggatcc ggccctctag atgcatgctc gagcggccgc cagtgtgatg gatatctgca
      361 gaattcgccc ttcaagatcg cgccactgta ctccagcctg ggtgacagag ggagaatctg
      421 tctcaaaaaa aagtttcttg acttctgttt ttggcagacc atgttctgcg aacacccagg
      481 agcactgctg agtgcaaaca tcatgtcgat tgcacaagtg gattcacgaa caggcgagga
      541 cgtgaccagg gacaaatgcg ggaaggggaa aggacgtgaa tcagaagcag agatgcttgc
      601 tgtcctacag caggcaggtg ggtgggtagg gtgcgagttc tgcggcccag gggttgggtt
      661 ttacactgaa gcaggacaga tgatgaagct aaggctccca gcaggtggga ggtggacggc
      721 gactcccatg tgaagcacag atgtgcaagg gatgcatcct cagtgaaagc gtggactagg
      781 aaaacagcaa cagcctggag actgttccta aactcagccc tcggcggaag ccaacccagg
      841 gtcattgcta gagggatcct cccccacctt gacaggagca aactcgaggg aagggcgaat
      901 tccagcacac tggcggtcga cttttttttt taagggcgaa ttctgcagat atccatcaca
      961 ctggcggccg ctcgagcatg catctagagg gcccaattcg ccctatagtg agtcgtatta
     1021 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1081 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     1141 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
     1201 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1261 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1321 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1381 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1441 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1501 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1561 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1621 attcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt ccgcagaaac
     1681 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg
     1741 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt
     1801 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc
     1861 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     1921 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     1981 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2041 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2101 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     2161 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2221 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatcc caccttgctc
     2281 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     2341 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2401 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     2461 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     2521 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     2581 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     2641 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     2701 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga attgaaaaag
     2761 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2821 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2881 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2941 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3001 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3061 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3121 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3181 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3241 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3301 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3361 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3421 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3481 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3541 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3601 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3661 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3721 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3781 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3841 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3901 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3961 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4021 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4081 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4141 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4201 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4261 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4321 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4381 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4441 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4501 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4561 agcggaag
//
