LOCUS       sequence-455929-        5884 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For subcloning of human GOLGA3 promoter or for assays using M13 
            phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5884)
  TITLE     Borchert Lab TOPO plasmids
REFERENCE   2  (bases 1 to 5884)
  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-GOLGA3
FEATURES             Location/Qualifiers
     source          1..5884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    99..120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(135..140,141..158,159..165)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    173..189
                     /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     178..200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     197..213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     197..213
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(279..290)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2309..2328)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2310..2328)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2335..2352)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2335..2351)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2344..2366)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2492..2920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2579..2598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2789..2810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3264..4058
                     /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(3318..3337)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3928..3947
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4076..4144,4145..4936)
                     /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(4294..4313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5107..5695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5596..5615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5849..5866
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
       61 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      121 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      181 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tggtaccgag
      241 ctcggatcca ctagtaacgg ccgccagtgt gctggaattc gcccttcaat acagcaacac
      301 acccagtctc ttaaaaaaaa aaaaaaaaaa tccatgagtc atctgtaaat ggcttacact
      361 ggcccagtga caataaactc caacctctta agaatccagc acctgcctac accaacatcc
      421 ttaccgccat cgctttcccc ctttgccatt acactgtggc cacatcctcc gccctcaggg
      481 tcttcaaact ttctgcctga ggcgcgcttt tctcacaatt ttgcctggct tgcttctcgt
      541 catagaggcc ttagcacgga agtcgcccgc ccccaggctc ccacttagag tagcggaccc
      601 cacccaacca ccgcccggta ccctctcgct ggcgaccctg ggtgtccctg tcctaggtgg
      661 gctccgcggg ggcagtgacc cagacccaca atacccagga cccgccacac gactgacatt
      721 cgtcgggttc atacccgtga caacgcccgg ccccactcgc tctggccacc tggaatgaag
      781 acaagtcggc cagcgacaaa gacaaacaag gaagtacacg gactcctgaa accccgactt
      841 cccacgaacc acccgacacc atgtcacgcc tttcagctgc aaagggccag gctaaagcaa
      901 gcactgcccg agtggtggca gtggacggct tcgggacagg ccgcgctcgt cgggcgcaga
      961 caagcattca ggagggctcg cgggcaccgg catttacaga gccagctccg ccgcagctga
     1021 aggcgggcgc ccaggcccga ggccaccgca gcgaggcggg ggtgactgga acggggccgt
     1081 cccgccacgc tgcagggacg cgcccaggcc ccgccgccgc ccctcgagcc tcccccgccc
     1141 atgcactgcg ggagcgggag cgggagcggg agcccgagcc ccgaggcgcc gcggcctccg
     1201 agcccctcac cctgctgctc tggcagcccc ggaggccgcc cggcccggat gctccggcgg
     1261 agacgtggcc gtgagagggg cggggcgagc ggtgcatcgg cctcgggtcc ccggaaacag
     1321 cctgcacgcg ctactttcgt tccggaacag agaacgtcct cctcttcttc aagtggccac
     1381 cctagggccg gggaccctct gcccacaggg tccggatcga gctcacctcc gactgtgatg
     1441 aggccgcctt cagaaagtga ccttaagagt ttgcggacct ttccctagta gaaacgcaac
     1501 ttccggaact gcaggaccaa ctctttcagg aagctgaacc gagcttccgg cggaagtggc
     1561 gtggccgggg ctcggcatgg gcgcggccga gctcggtgtg ggcgtgactt ccccgcccgc
     1621 cctagtcggg tccccgcccc aaccccgcac ctccacccgg ccggcgccct tcgcgcttca
     1681 ggtgccagat caaaggcgac gctgctttgg accctatttg cttccccttt aaaaagggca
     1741 aagctggacg gcgcgggccg cgggccgctg gggaacggct ttccaccttc cctcatccgc
     1801 caggccgctc ttccagctgt caggaaggaa atggcctgga gggcgcctcg gggtcctctg
     1861 tgatgttgcg ctcatctctg cggattttgg aatacgcttc agtctacgaa gttttctggg
     1921 attttgatgg gaactgctgc agatcgtttg gggagcacgg gcatcgctga ctcggtcctt
     1981 catctgcgtc cctggtggcc cctccattta tttcggtctt ttgatttctt ccaccagcat
     2041 tttgtaattt cagcatacag attctgcacc ttttaagttt atagtatttc acatcaaaca
     2101 actgtaacac tgcgtttttg acgttttcca cgtgttcact gttggtctac acaaatgtga
     2161 tgaattctat gttgattttc ttatcctgag acctcactga actcattagt ttgagggttc
     2221 atttggtgtt ttgcggttga agggcgaatt ctgcagatat ccatcacact ggcggccgct
     2281 cgagcatgca tctagagggc ccaattcgcc ctatagtgag tcgtattaca attcactggc
     2341 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     2401 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     2461 ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg
     2521 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2581 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2641 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2701 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2761 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2821 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2881 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat tcagggcgca
     2941 agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc
     3001 ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc
     3061 aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa
     3121 gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa
     3181 actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag
     3241 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3301 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3361 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3421 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3481 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3541 tattgggcga agtgccgggg caggatctcc tgtcatccca ccttgctcct gccgagaaag
     3601 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3661 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3721 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3781 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3841 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3901 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3961 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4021 gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat tgaaaaagga agagtatgag
     4081 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4141 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4201 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4261 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4321 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4381 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4441 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4501 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4561 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4621 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4681 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4741 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4801 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4861 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4921 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4981 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5101 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5161 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5221 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5281 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5341 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5401 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5461 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5521 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5581 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5641 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5701 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5761 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5821 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5881 ccca
//
