LOCUS       sequence-499133-        4486 bp    DNA     circular SYN 04-FEB-2026
DEFINITION  Cell-free translocation reporter; proOmpA (1-178); C-terminal pep99 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4486)
  AUTHORS   Meier M, Scholz SA, von Bank L, Levandoski JE, Luckhof M, Schaaf M, 
            Si S, Lieberwirth I, Jung AL, Landfester K, Driessen AJM, Erb TJ
  TITLE     Functional Mapping and Engineering of the Sec Translocon Unlocked by
            a Cell-Free System
  JOURNAL   bioRxiv 2025.12.09.688994
REFERENCE   2  (bases 1 to 4486)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.12.09.688994"
COMMENT     Sequence Label: pTE5602
FEATURES             Location/Qualifiers
     source          1..4486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(548..1363)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(548..1363)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1271..1290
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1355..1374
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1485..2073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1974..1993
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2227..2244
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2259..2398
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2324..2343)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2484..2506
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2500..2691)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3540..3559
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     sig_peptide     3690..3752
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     primer_bind     complement(4389..4407)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4403..4450
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa
      541 ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
      601 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
      661 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
      721 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
      781 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
      841 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
      901 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
      961 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     1021 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     1081 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     1141 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     1201 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     1261 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     1321 atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     1381 tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg
     1441 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1501 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1561 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1621 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1681 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1741 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1801 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1861 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1921 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1981 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2041 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2101 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2161 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2221 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     2281 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     2341 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     2401 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2461 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2521 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     2581 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     2641 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctaatgcctc
     2701 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     2761 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     2821 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     2881 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     2941 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     3001 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     3061 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     3121 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     3181 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     3241 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     3301 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     3361 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     3421 tgggttggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     3481 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     3541 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     3601 atgcgtccgg cgtagaggat cgagatctcg ggtaatacga ctcactatag atactcgcta
     3661 gggcgaatac tattataagg aggactttta tgaaaaagac agctatcgcg attgcagtgg
     3721 cactggctgg tttcgctacc gtagcgcagg ccgctccgaa agataacacc tggtacactg
     3781 gtgctaaact gggctggtcc cagtaccatg acactggttt catcaacaac aatggcccga
     3841 cccatgaaaa ccaactgggc gctggtgctt ttggtggtta ccaggttaac ccgtatgttg
     3901 gctttgaaat gggttacgac tggttaggtc gtatgccgta caaaggcagc gttgaaaacg
     3961 gtgcatacaa agctcagggc gttcaactga ccgctaaact gggttaccca atcactgacg
     4021 acctggacat ctacactcgt ctgggtggca tggtatggcg tgcagacact aaatccaacg
     4081 tttatggtaa aaaccacgac accggcgttt ctccggtctt cgctggcggt gttgagtacg
     4141 cgatcactcc tgaaatcgct acccgtctgg aataccagtg gaccaacaac atcggtgacg
     4201 cacacaccat cggcactcgt ccgggaggct ccggcgtgac cggctaccgc ctgtttgaaa
     4261 aaattagcta acctcggtac caaattccag aaaagaggcc tcccgaaagg ggggcctttt
     4321 ttcgttttgg tcccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     4381 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     4441 gggttttttg ctgaaaggag gaactatatc cggattggcg aatggg
//
