LOCUS       sequence-387064-        7144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the two Strep/one FLAG (TSF)-tagged human tepsin in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7144)
  AUTHORS   Mattera R, Guardia CM, Sidhu SS, Bonifacino JS
  TITLE     Bivalent Motif-Ear Interactions Mediate the Association of the 
            Accessory Protein Tepsin with the AP-4 Adaptor Complex.
  JOURNAL   J Biol Chem. 2015 Dec 25;290(52):30736-49. doi: 
            10.1074/jbc.M115.683409. Epub 2015 Nov 5.
  PUBMED    26542808
REFERENCE   2  (bases 1 to 7144)
  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"; doi: 
            "10.1074/jbc.M115.683409"; journalName: "J Biol Chem"; date: 
            "2015-12-25- 25"; volume: "290"; issue: "52"; pages: "30736-49"
COMMENT     Sequence Label: pcDNA 3.1-TSF-tepsin
FEATURES             Location/Qualifiers
     source          1..7144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      169..304
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             385..1179
                     /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(439..458)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1049..1068
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1355..1476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1392..1411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1446..1465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1525..1541)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1525..1541)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1538..1560)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1549..1565
                     /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(1573..1579,1580..1597,1598..1603))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1618..1639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1756..1773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1927..2515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2007..2026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2686..3477,3478..3546))
                     /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     3309..3328
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3547..3651)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3726..3745)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3917..4296
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4297..4500
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4451..4471
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4545..4564
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4545..4563
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4596..4605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4611..4634
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4671..4694
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4701..4724
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6330..6359
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6375..6392
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6415..6432)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6421..6645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6691..7119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6778..6797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6988..7009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(join(7128..7144,1..4))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(7133..7144,1..318)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
       61 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
      121 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
      181 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
      241 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
      301 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
      361 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
      421 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
      481 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
      541 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
      601 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
      661 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
      721 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
      781 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
      841 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
      901 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
      961 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     1021 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     1081 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     1141 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     1201 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     1261 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     1321 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     1381 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1441 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     1501 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1561 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1621 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1681 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1741 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1801 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1861 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2401 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2461 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2521 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2581 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2641 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2701 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2761 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2821 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2881 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2941 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3001 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3061 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3121 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3181 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3241 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3301 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3361 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3421 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3481 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3541 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3601 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3661 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     3721 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     3781 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     3841 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     3901 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     3961 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     4021 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     4081 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     4141 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     4201 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     4261 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     4321 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     4381 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     4441 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     4501 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
     4561 agggagaccc aagctggcta gttaagctta tcgatgccac catggctagc tggagccacc
     4621 cgcagttcga gaaaggtgga ggtgcccgag gtggatcggg aggtggatcg tggagccacc
     4681 cgcagttcga aaaaggattc gattacaagg atgacgatga caagggtacc atggctgccg
     4741 cgccgccgct acgggaccgc ctgagctttc tacaccggct cccgattctc ctgaagggga
     4801 cgtccgatga tgatgtcccg tgtccgggct acctgtttga agagattgct aaaatctccc
     4861 acgagtctcc gggcagcagc cagtgcctgc tggagtacct cctgagccgc ctgcacagca
     4921 gctccggcca cgggaagctc aaggtgctga agatcctgct ctatctgtgc agccacggct
     4981 cctccttctt cctgctcatc ctcaaacgca actctgcctt catccaggaa gctgcagctt
     5041 ttgcagggcc cccagatcct ctgcacggga acagcttgta ccagaaggtt cgcgcggccg
     5101 cgcaggactt ggggagcacc ctgttctcgg acaccgtgtt gccgctggct ccctcccagc
     5161 ctctggggac cccgcctgcc acaggcatgg gctcccaggc caggccgcac agcaccctcc
     5221 agggtttcgg ctacagcaag gaacacggcc gcacggctgt gaggcatcag cctgggcagg
     5281 ccggaggggg ctgggatgag ctggacagcg gccccagctc tcagaattcc tcccagaaca
     5341 gtgacctgag cagggtctcg gactcgggca gtcattccgg cagcgacagc cattcagggg
     5401 ccagccggga gccgggtgac ctggcagaaa gggtcgaggt ggtggccctg agtgactgtc
     5461 agcaggagtt gagcttggtg aggactgtga ctcggggacc acgcgccttc ctgagccgcg
     5521 aggaggcaca gcacttcatc aaagcgtgtg gactgctcaa ctgtgaggcc gtgctgcagc
     5581 tgctgacctg ccacctgcgt gggaccagtg aatgcacgca gctgagggcg ctgtgtgcca
     5641 tcgcctccct ggggagcagc gacctcctcc cccaggagca catcctcctc cgcacccggc
     5701 cgtggctgca ggagctcagc atgggcagcc cgggacctgt gaccaacaag gccaccaaga
     5761 tcctgaggca ctttgaggcc tcctgtgggc agctgtcccc tgcccggggc acctcagctg
     5821 agcctggccc cacagccgcc ctcccaggcc catctgacct gctgaccgac gctgtgcctc
     5881 tccctgggag ccaggtcttc ctgcagcctc tgagttcaac cccggtctcg tcccggagcc
     5941 ctgctccctc atctgggatg ccgtccagcc ctgtgcccac cccaccccca gatgcctccc
     6001 ccattccagc ccccggagac cccagcgagg ccgaggccag actggcagaa agcaggcggt
     6061 ggagacctga acggatccca gggggcacgg acagcccaaa gagaggcccc agcagctgtg
     6121 cgtggagccg cgactccttg tttgctggca tggagctggt ggcctgtccc cgcctggtgg
     6181 gggctggggc tgctgcggga gagtcctgtc ctgatgctcc ccgcgccccc caaacatcgt
     6241 cccagaggac agcagccaaa gagcctcctg gctcagagcc gtcagctttc gcgttcctga
     6301 acgcctgact cgagtctaga gggcccttcg aacaaaaact catctcagaa gaggatctga
     6361 atatgcatac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     6421 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     6481 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     6541 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     6601 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     6661 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     6721 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     6781 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     6841 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     6901 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     6961 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     7021 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     7081 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     7141 gtta
//
