LOCUS       sequence-442808-        6731 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 6731)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 6731)
  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_miniTurbo_Cterm
FEATURES             Location/Qualifiers
     source          1..6731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    987..1011
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1795..1819)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1839..2609
                     /codon_start=1
                     /product="N-terminally truncated mutant of the E. coli 
                     biotin ligase BioID engineered for improved catalytic 
                     efficiency (Branon et al., 2018)"
                     /label=miniTurbo
                     /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             2619..2642
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2683..2700)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2689..2913
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3046..3065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3197..3244
                     /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             3252..4274
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4404..4525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4441..4460)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4495..4514
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4574..4590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4574..4590)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4587..4609)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4598..4614
                     /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(4622..4628,4629..4646,4647..4652))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4667..4688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4805..4822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4976..5564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5056..5075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5735..6526,6527..6595))
                     /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"
     promoter        complement(6596..6700)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag ctttagacaa gtttgtacaa aaaagttggc atgtctcgaa
     1021 gatatgactc caggaccact atattttctc cagaaggtcg cttataccaa gttgaatatg
     1081 ccatggaagc tattggacat gcaggcacct gtttgggaat tttagcaaat gatggtgttt
     1141 tgcttgcagc agagagacgc aacatccaca agcttcttga tgaagtcttt ttttctgaaa
     1201 aaatttataa actcaatgag gacatggctt gcagtgtggc aggcataact tctgatgcta
     1261 atgttctgac taatgaacta aggctcattg ctcaaaggta tttattacag tatcaggagc
     1321 caataccttg tgagcagttg gttacagcgc tgtgtgatat caaacaagct tatacacaat
     1381 ttggaggaaa acgtcccttt ggtgtttcat tgctgtacat tggctgggat aagcactatg
     1441 gctttcagct ctatcagagt gaccctagtg gaaattacgg gggatggaag gccacatgca
     1501 ttggaaataa tagcgctgca gctgtgtcaa tgttgaaaca agactataaa gaaggagaaa
     1561 tgaccttgaa gtcagcactt gctttagcta tcaaagtact aaataagacc atggatgtta
     1621 gtaaactctc tgctgaaaaa gtggaaattg caacactaac aagagagaat ggaaagacag
     1681 taatcagagt tctcaaacaa aaagaagtgg agcagttgat caaaaaacac gaggaagaag
     1741 aagccaaagc tgagcgtgag aagaaagaaa aagaacagaa agaaaaggat aaatacccaa
     1801 ctttcttgta caaagtggtt gattctagaa tggctagcat cccgctgctg aacgctaaac
     1861 agattctggg acagctggac ggcgggagcg tggcagtcct gcctgtggtc gactccacca
     1921 atcagtacct gctggatcga atcggcgagc tgaagagtgg ggatgcttgc attgcagaat
     1981 atcagcaggc agggagagga agcagaggga ggaaatggtt ctctcctttt ggagctaacc
     2041 tgtacctgag tatgttttgg cgcctgaagc ggggaccagc agcaatcggc ctgggcccgg
     2101 tcatcggaat tgtcatggca gaagcgctgc gaaagctggg agcagacaag gtgcgagtca
     2161 aatggcccaa tgacctgtat ctgcaggata gaaagctggc aggcatcctg gtggagctgg
     2221 ccggaataac aggcgatgct gcacagatcg tcattggcgc cgggattaac gtggctatga
     2281 ggcgcgtgga ggaaagcgtg gtcaatcagg gctggatcac actgcaggaa gcagggatta
     2341 acctggacag gaatactctg gccgctatgc tgatccgaga gctgcgggca gccctggaac
     2401 tgttcgagca ggaaggcctg gctccatatc tgtcacggtg ggagaagctg gataacttca
     2461 tcaatagacc cgtgaagctg atcattgggg acaaagagat tttcgggatt agccggggga
     2521 ttgataaaca gggagccctg ctgctggaac aggacggagt tatcaaaccc tggatgggcg
     2581 gagaaatcag tctgcggtct gccgaaaagc tgcaggacga ctacaaagac gatgacgata
     2641 aataactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     2701 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2761 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2821 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2881 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2941 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3001 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3061 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3121 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3181 atggttcacg tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa
     3241 cttccttggc caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     3301 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     3361 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     3421 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     3481 ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     3541 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     3601 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     3661 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     3721 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     3781 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     3841 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     3901 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     3961 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     4021 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     4081 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     4141 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     4201 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     4261 gggcaaagga atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     4321 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4381 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4441 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4501 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4561 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4621 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4681 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4741 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4801 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4861 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4921 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4981 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5041 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5101 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5161 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5221 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5281 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5341 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5401 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5461 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5521 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5581 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5641 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5701 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5761 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5821 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5881 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5941 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6001 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6061 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6121 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6181 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6241 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6301 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6361 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6421 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6481 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6541 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6601 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6661 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6721 cacctgacgt c
//
