LOCUS       sequence-319720-        6122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for EF1n3_mFAP2b 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 6122)
  TITLE     Baker Lab mFAP plasmids
REFERENCE   2  (bases 1 to 6122)
  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-EF1n3_mFAP2b
FEATURES             Location/Qualifiers
     source          1..6122
                     /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(624..642)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      638..685
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(860..865,866..882,883..888))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(915..933)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1001..1105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1106..1174,1175..1966)
                     /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(1324..1343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2137..2725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2626..2645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2879..2896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2911..3050
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2976..2995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3136..3158
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3152..3343)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5698..5717
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5737..5814)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6020..6039
                     /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 gtatgagccg tgcggcgcag ctcctgccgg gtacctggca ggtgaccatg
      241 accaacgaag acggtcagac ctcccagggc cagtggcact tccagccgcg ttctccgtac
      301 accatggaca ttgttgctca gggtaccatc tctgacggtc gtccgatcgt tggttacggt
      361 aaagcgaccg ttaaaacccc ggacaccctg gacatagaca tcacctaccc gtctctgggt
      421 aacatcaaag ctcagggtca gataaccatg gactctccga cccagttcaa atgggacgca
      481 catgtggaca aagatggtga cggctatatt tccgccgccg aagcagcagc tcagccgcgt
      541 ctgaccggta ccctgcagcg tcaggaataa tgaggatccg gctgctaaca aagcccgaaa
      601 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
      661 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag
      721 aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg
      781 aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt
      841 aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac atgagaattc
      901 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
      961 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1021 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1081 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1141 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1201 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1261 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1321 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     1381 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1441 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1501 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1561 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1621 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     1681 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1741 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1801 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     1861 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1921 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1981 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2041 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2101 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2161 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2221 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2281 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2341 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2401 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2461 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2521 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2581 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2641 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2701 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2761 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2821 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2881 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     2941 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3001 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     3061 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3121 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3181 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     3241 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     3301 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     3361 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     3421 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     3481 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
     3541 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
     3601 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     3661 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     3721 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     3781 acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc
     3841 gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg
     3901 cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag
     3961 cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa
     4021 cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt
     4081 gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct
     4141 ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga
     4201 cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg
     4261 gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc
     4321 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     4381 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     4441 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     4501 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     4561 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     4621 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4681 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4741 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     4801 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     4861 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     4921 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     4981 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     5041 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     5101 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     5161 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     5221 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     5281 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     5341 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     5401 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     5461 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     5521 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     5581 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5641 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     5701 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     5761 ttccgggcgc tatcatacca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     5821 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     5881 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     5941 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     6001 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     6061 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
     6121 at
//
