LOCUS       sequence-316643-        6095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for mFAP10 with an N-terminal 6xHis tag, thrombin epitope, 
            and TEV epitope. Expression is controlled by the T7 promoter with E.
            coli codon optimization..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6095)
  TITLE     Baker Lab mFAP plasmids
REFERENCE   2  (bases 1 to 6095)
  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: pET15-mFAP10
FEATURES             Location/Qualifiers
     source          1..6095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             166..186
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(597..615)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      611..658
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(833..838,839..855,856..861))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(888..906)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        974..1078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1079..1147,1148..1939)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1297..1316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2110..2698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2599..2618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2852..2869
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2884..3023
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2949..2968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3109..3131
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3125..3316)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5671..5690
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5710..5787)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5993..6012
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatactatg gctagcagcc atcatcatca tcatcatagc
      121 agcggcctgg tgccgcgcgg cagctccatg agccgtgcga gcggcgagaa cctgtacttt
      181 cagggttcga gtatgtcacg cgccgcgcag ctcctgccag gtacctggca ggcaaccttt
      241 accaacgaag acggtcagac ctctcaaggt cagattcact tccagccgcg ttctccgtac
      301 accatggaca ttgttgctca gggtaccatc tctgacggtc gtccgatcgt tggttacggt
      361 aaagcgaccg ttaaaacccc ggacaccctg gacatagaca tcacctaccc gtctctgggt
      421 aacatcaaag ctcagggtca gataaccatg gactctccga cccagttcaa atttgacgca
      481 accaccaaag gtgaaaacga tttccatggt cgtctgaccg gtaccctgca gcgtcaggaa
      541 taatgaggat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
      601 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      661 gaaaggagga actatatccg gatatcccac aagaggcccg gcagtaccgg cataaccaag
      721 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
      781 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
      841 ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg
      901 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
      961 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1021 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1081 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1141 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1201 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1261 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1321 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1381 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1441 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1501 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1561 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1621 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1681 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1741 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1801 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1861 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1921 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1981 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2041 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2101 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2161 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2221 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2281 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2341 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2401 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2461 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2521 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2581 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2641 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2701 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2761 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2821 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2881 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     2941 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     3001 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     3061 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3121 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3181 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3241 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3301 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     3361 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     3421 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     3481 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     3541 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     3601 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     3661 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     3721 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     3781 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
     3841 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
     3901 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
     3961 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
     4021 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
     4081 gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
     4141 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
     4201 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
     4261 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     4321 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     4381 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     4441 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     4501 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4561 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     4621 gttgcgctca ctgcccgctt tccagtcggg aaactgtcgt gccagctgca ttaatgaatc
     4681 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     4741 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     4801 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     4861 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     4921 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     4981 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     5041 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     5101 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     5161 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     5221 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     5281 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     5341 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     5401 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     5461 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     5521 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     5581 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     5641 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     5701 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     5761 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     5821 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     5881 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     5941 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     6001 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     6061 cggcgtagag gatcgagatc tcgatcccgc gaaat
//
