LOCUS       sequence-439223-        6935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag-tagged biotinylation enzyme for co-expression of Psma4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6935)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 6935)
  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: PSMA4_BirA_Cterm
FEATURES             Location/Qualifiers
     source          1..6935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(3..794,795..863))
                     /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     626..645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(864..968)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1043..1062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1234..1613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1614..1817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1768..1788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1819..1837
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1840..1858
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1868..1892
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1986..2010
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2794..2818)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2829..3179,3180..3182,3183..3791)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             3822..3845
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3886..3903)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3892..4116
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4249..4268)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4400..4447
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4455..5477
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5607..5728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5644..5663)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5698..5717
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5777..5793)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5777..5793)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5790..5812)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5801..5817
                     /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(5825..5831,5832..5849,5850..5855))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5870..5891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6008..6025)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6179..6767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6259..6278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
       61 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
      121 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
      181 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
      241 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
      301 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
      361 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
      421 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
      481 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
      541 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
      601 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
      661 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
      721 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
      781 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
      841 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
      901 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
      961 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     1021 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     1081 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     1141 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     1201 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     1261 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     1321 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     1381 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     1441 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     1501 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     1561 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     1621 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     1681 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     1741 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     1801 tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt gatagagatc
     1861 gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     1921 acctccatag aagacaccgg gaccgatcca gcctccggac tctagcgttt aaacttaagc
     1981 tttagacaag tttgtacaaa aaagttggca tgtctcgaag atatgactcc aggaccacta
     2041 tattttctcc agaaggtcgc ttataccaag ttgaatatgc catggaagct attggacatg
     2101 caggcacctg tttgggaatt ttagcaaatg atggtgtttt gcttgcagca gagagacgca
     2161 acatccacaa gcttcttgat gaagtctttt tttctgaaaa aatttataaa ctcaatgagg
     2221 acatggcttg cagtgtggca ggcataactt ctgatgctaa tgttctgact aatgaactaa
     2281 ggctcattgc tcaaaggtat ttattacagt atcaggagcc aataccttgt gagcagttgg
     2341 ttacagcgct gtgtgatatc aaacaagctt atacacaatt tggaggaaaa cgtccctttg
     2401 gtgtttcatt gctgtacatt ggctgggata agcactatgg ctttcagctc tatcagagtg
     2461 accctagtgg aaattacggg ggatggaagg ccacatgcat tggaaataat agcgctgcag
     2521 ctgtgtcaat gttgaaacaa gactataaag aaggagaaat gaccttgaag tcagcacttg
     2581 ctttagctat caaagtacta aataagacca tggatgttag taaactctct gctgaaaaag
     2641 tggaaattgc aacactaaca agagagaatg gaaagacagt aatcagagtt ctcaaacaaa
     2701 aagaagtgga gcagttgatc aaaaaacacg aggaagaaga agccaaagct gagcgtgaga
     2761 agaaagaaaa agaacagaaa gaaaaggata aatacccaac tttcttgtac aaagtggttg
     2821 attctagaat gaaggacaac accgtgcccc tgaagctgat cgccctgctg gccaacggcg
     2881 agttccacag cggcgagcag ctgggcgaga ccctgggcat gagccgggcc gccatcaaca
     2941 agcacatcca gaccctgcgg gactggggcg tggacgtgtt caccgtgccc ggcaagggct
     3001 acagcctgcc cgagcccatc cagctgctga acgccaagca gatcctgggc cagctggacg
     3061 gcggcagcgt ggccgtgctg cccgtgatcg acagcaccaa ccagtacctg ctggaccgga
     3121 tcggcgagct gaagagcggc gacgcctgca tcgccgagta ccagcaggcc ggccggggcg
     3181 gccggggccg gaagtggttc agccccttcg gcgccaacct gtacctgagc atgttctggc
     3241 ggctggagca gggccccgcc gccgccatcg gcctgagcct ggtgatcggc atcgtgatgg
     3301 ccgaggtgct gcggaagctg ggcgccgaca aggtgcgggt gaagtggccc aacgacctgt
     3361 acctgcagga ccggaagctg gccggcatcc tggtggagct gaccggcaag accggcgacg
     3421 ccgcccagat cgtgatcggc gccggcatca acatggccat gcggcgggtg gaggagagcg
     3481 tggtgaacca gggctggatc accctgcagg aggccggcat caacctggac cggaacaccc
     3541 tggccgccat gctgatccgg gagctgcggg ccgccctgga gctgttcgag caggagggcc
     3601 tggcccccta cctgagccgg tgggagaagc tggacaactt catcaaccgg cccgtgaagc
     3661 tgatcatcgg cgacaaggag atcttcggca tcagccgggg catcgacaag cagggcgccc
     3721 tgctgctgga gcaggacggc atcatcaagc cctggatggg cggcgagatc agcctgcgga
     3781 gcgccgagaa ggggggatct ggccccggcg gcggcgcgcc agactacaaa gacgatgacg
     3841 ataaataact cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     3901 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3961 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4021 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4081 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4141 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4201 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4261 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4321 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4381 gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag
     4441 gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     4501 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     4561 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     4621 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     4681 tgcttgacat tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     4741 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     4801 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     4861 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     4921 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     4981 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     5041 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     5101 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     5161 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     5221 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     5281 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     5341 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     5401 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     5461 cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa
     5521 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5581 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5641 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5701 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5761 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5821 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5881 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5941 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6001 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6061 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6121 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6181 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6241 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6301 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6361 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6421 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6481 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6541 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6601 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6661 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6721 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6781 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6841 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6901 caatctaaag tatatatgag taaacttggt ctgac
//
