LOCUS       sequence-421141-        6170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6170)
  TITLE     Yaffe lab HA-tagged 14-3-3 plasmids
REFERENCE   2  (bases 1 to 6170)
  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"
COMMENT     Sequence Label: pcDNA3-HA-14-3-3 gamma
FEATURES             Location/Qualifiers
     source          1..6170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3..23)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8..337
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      188..323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     250..269
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             404..1198
                     /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(458..477)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1068..1087
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1372..1493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1409..1428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1463..1482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1542..1558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1542..1558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1555..1577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1566..1582
                     /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(1590..1596,1597..1614,1615..1620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1635..1656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1773..1790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1944..2532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2024..2043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2703..3494,3495..3563))
                     /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     3326..3345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3564..3668)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3743..3762)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3934..4313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4314..4517
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4468..4488
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4562..4581
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4562..4580
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4603..4629
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             4603..4629
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4609..4635
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(5422..5440)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5423..5440)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5460..5477)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5466..5690
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5736..6164
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5823..5842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6033..6054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
       61 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
      121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
      181 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
      241 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
      301 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
      361 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
      421 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
      481 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
      541 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
      601 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
      661 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
      721 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
      781 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
      841 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
      901 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
      961 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     1021 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     1081 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     1141 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     1201 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     1261 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     1321 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     1381 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1441 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     1501 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     1561 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1621 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1681 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1741 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1801 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1861 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1921 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1981 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2041 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2101 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2161 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2221 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2281 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2341 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2401 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2461 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2521 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2581 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2641 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2701 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2761 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2821 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2881 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2941 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3001 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3061 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3121 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3181 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3241 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3301 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3361 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3421 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3481 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3541 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3601 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3661 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     3721 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     3781 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     3841 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     3901 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     3961 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4021 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4081 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4141 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4201 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4261 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     4321 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4381 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4441 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4501 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
     4561 ttaatacgac tcactatagg gagacccaag cttaccatgg cctaccccta cgacgtgccc
     4621 gactacgcct ccctcggatc cccggaattc atggtggacc gcgagcaact agtgcagaaa
     4681 gcccggctgg ccgagcaggc ggagcgctac gacgatatgg ccgcggccat gaagaacgtg
     4741 acagagctga atgaaccact gtccaacgag gaacggaacc tcctgtcggt ggcctacaag
     4801 aacgtggttg gggctcgccg ctcctcctgg agggtcatca gcagcatcga gcagaagacg
     4861 tctgcggatg gcaacgagaa gaagatagag atggtccgag cctaccggga aaagattgag
     4921 aaggagctgg aggccgtgtg ccaggacgtg ctgagcctgc tggacaacta cctgatcaag
     4981 aactgcagcg agacccagta cgagagcaag gtgttctacc tgaagatgaa aggggactat
     5041 taccgctacc tggctgaagt ggccactggg gagaaaaggg cgaccgtggt ggagtcgtcc
     5101 gagaaggcct acagcgaagc ccacgagatc agcaaggagc acatgcagcc cacccacccc
     5161 atccggctgg gcctggcgct caactactcc gttttctact acgagatcca gaacgcccca
     5221 gagcaagcct gccacctggc caagaccgcc ttcgatgacg ccatcgccga gctcgacact
     5281 ctgaacgagg actcctacaa ggactccact ctgatcatgc agctgctccg agacaaccta
     5341 acgctctgga cgagcgacca gcaagacgac gacggcggcg aaggcaacaa ctaactcgag
     5401 catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc
     5461 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     5521 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     5581 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     5641 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     5701 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     5761 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5821 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5881 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5941 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6001 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6061 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6121 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
//
