LOCUS       sequence-420484-        6399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of the N-terminal amino acids 1-175 of human 
            C17orf53 N-terminally fused to Profinity eXact-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6399)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 6399)
  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: pPAL7-C17orf53_1-175
FEATURES             Location/Qualifiers
     source          1..6399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15..33
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    34..58
                     /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             73..95
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(914..932)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      928..975
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1150..1155,1156..1172,1173..1178))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1205..1223)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1291..1395
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1396..1464,1465..2256)
                     /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(1614..1633)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2427..3015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2916..2935
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3169..3186
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3201..3340
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3266..3285)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3426..3448
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3442..3633)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4945..6027)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5989..6008
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6028..6105)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6311..6330
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
       61 tctagaaata attttgttta actttaagaa ggagatatac atatgggagg gaaatcaaac
      121 ggggaaaaga aatatattgt cgggttcaaa cagggcttta agagctgcgc taagaaggag
      181 gatgtcattt ctgaaaaagg cgggaaactc caaaagtgct tcaaatatgt agacgcagct
      241 agcgctacat taaacgaaaa agctgtagaa gaattgaaaa aagatccgag cgtcgcgtac
      301 gtagaagaag acaagctctt caaagctttg atggcgtgca gtttgcagaa gctgtttgct
      361 gtggaagagg agtttgaaga tgaggatttc ttgtctgctg tggaggatgc agagaaccgg
      421 tttactggct cactgcctgt gaatgctggg cgcctgagac ctgtctcttc taggccacag
      481 gagactgtgc aggcacagtc ctccaggctg ctgctgttac accccactgc tccctcagag
      541 gctttgggcc tgccagactt ggacctctgc ctccctgcct ccagcacgcc cagtgctgac
      601 agccgtccat catgcatagg agcagctccc ctaaggcctg tctctacttc cagcagctgg
      661 attggcaatc agagaagagt gacagtgaca gaagtgctca gagagacagc aagacctcag
      721 tcctcagcct tacaccccct actcaccttt gagagccaac agcagcaagt tggtggcttt
      781 gaggggcctg aacaagacga atttgataaa gtcctggcaa gcatggagtt ggaggagcct
      841 ggcatggagc tggaatgaga attcctcgag gcggccgcat aagcccgaaa ggaagctgag
      901 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
      961 ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca
     1021 gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga
     1081 tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata
     1141 aactaccgca ttaaagccta tcgatgataa gctgtcaaac atgagatttc ttgaagacga
     1201 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1261 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1321 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1381 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1441 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1501 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1561 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1621 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     1681 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1741 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1801 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1861 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1921 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1981 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2041 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2101 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2161 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2221 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2281 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2341 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2401 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2461 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2521 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2581 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2641 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2701 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2761 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2821 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2881 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2941 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3001 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3061 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3121 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3181 agcgaggaag cggaacagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3241 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     3301 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
     3361 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3421 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     3481 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     3541 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     3601 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     3661 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     3721 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     3781 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     3841 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     3901 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     3961 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     4021 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     4081 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct
     4141 gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt
     4201 tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg
     4261 ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg
     4321 cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag
     4381 gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc
     4441 gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc
     4501 tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc
     4561 cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg
     4621 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     4681 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     4741 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     4801 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     4861 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     4921 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4981 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     5041 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     5101 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     5161 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     5221 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     5281 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     5341 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     5401 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     5461 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     5521 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     5581 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     5641 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     5701 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     5761 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     5821 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     5881 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     5941 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     6001 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     6061 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     6121 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     6181 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     6241 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     6301 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     6361 tgccggccac gatgcgtccg gcgtagagga tcgagatct
//
