LOCUS       sequence-335414-        6769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FepA D34C, delta12-16 (TonB box) . Please note that mutation
            numbers are not based on start codon of insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6769)
  AUTHORS   Postle K, Kho K, Gresock M, Ghosh J, Larsen R
  TITLE     Identification of New In Vivo TonB-FepA Rendezvous Sites
  JOURNAL   bioRxiv 2021.12.08.471779
REFERENCE   2  (bases 1 to 6769)
  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"; journalName: "bioRxiv 
            2021.12.08.471779"
COMMENT     Sequence Label: pKP1626
FEATURES             Location/Qualifiers
     source          1..6769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(2627..2644)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2797..2883
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2975..3002
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3021..3112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3113..3181,3182..3973)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3331..3350)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4015..4470
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4102..4121)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4312..4333
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4581..5169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5070..5089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5323..5340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5506..5528
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5666..5685
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(5865..6743)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagcaggag gaattcacca tgaacaagaa gattcattcc ctggccttgt tggtcaatct
      361 ggggatttat ggggtagcgc aggcacaaga gccgacggat actcctgttt cacatgacac
      421 cgccgccgag cagaacttac aggcgcctgg cgtttcgacc atcaccgcat gcgaaatccg
      481 caaaaacccg gttgcccgcg atgtgtcgaa gatcatccgt accatgccag gcgttaacct
      541 gaccggtaac tccaccagtg gtcagcgtgg gaataaccga cagattgata ttcgcggtat
      601 gggtccggaa aacacgctga ttttgattga cggcaagccg gtaagcagcc gtaactcggt
      661 gcgtcagggc tggcgtggcg agcgcgatac ccgtggtgat acttcctggg tgccacctga
      721 aatgattgaa cgtattgaag ttctgcgtgg tccggcagct gcgcgttatg gcaacggcgc
      781 ggcgggcggc gtggttaaca tcattaccaa aaaaggcagc ggcgagtggc acggctcctg
      841 ggacgcatat ttcaatgcgc cagaacataa agaggaaggt gccaccaaac gcactaactt
      901 tagcctgacc ggtccgctgg gcgacgaatt cagcttccgt ttgtatggca acctcgacaa
      961 aacccaggct gacgcgtggg atatcaacca gggccatcag tccgcgcgtg ccggaacgta
     1021 tgccacgacg ttaccagccg ggcgcgaagg ggtaatcaac aaagatatta atggcgtggt
     1081 gcgctgggat ttcgcgccat tgcaatcgct ggaactggaa gcaggttaca gccgccaggg
     1141 taacctgtat gcgggcgaca cccagaatac caactccgat tcctataccc gctcgaaata
     1201 tggcgatgaa accaaccgtc tgtatcgcca gaactacgcg ctgacctgga acggtggctg
     1261 ggataacggc gtgaccacca gcaactgggt gcagtacgaa cacacccgta actcgcgtat
     1321 tccggaaggt ctggcgggcg gtaccgaagg gaaatttaac gaaaaagcga cacaggattt
     1381 cgtcgatatc gatcttgatg acgtgatgct gcacagcgaa gttaacctgc cgattgattt
     1441 cctcgttaac cagacgctga cgctgggtac ggagtggaat cagcaacgga tgaaggactt
     1501 aagttccaac acccaggcac tgaccggaac gaataccggt ggcgctattg atggcgtgag
     1561 taccaccgac cgtagcccgt attcaaaagc agaaattttc tcgctgtttg ccgaaaacaa
     1621 catggagctg actgacagca ccatcgtaac gccggggctg cgtttcgatc atcacagtat
     1681 tgtcggcaat aactggagcc cggcgctgaa catatcgcaa ggtttaggcg atgacttcac
     1741 gctgaaaatg ggcatcgccc gtgcttataa agcgccgagc ctgtaccaga ctaacccgaa
     1801 ctacattctc tacagtaaag gtcagggttg ctatgccagc gcgggcggct gctatctgca
     1861 aggtaacgat gacctgaaag cagaaaccag catcaacaaa gagattggtc tggagttcaa
     1921 acgcgacggg tggctggcgg gcgtcacctg gttccgtaac gattatcgca ataagattga
     1981 agcgggctat gtggctgtag ggcaaaacgc agtcggcacc gatctctatc agtgggataa
     2041 cgtgccgaaa gcggtggttg aaggtctgga aggatcgtta aacgtaccgg ttagcgaaac
     2101 ggtgatgtgg accaataaca tcacttatat gctgaagagt gaaaacaaaa ccacgggcga
     2161 ccgtttgtcg atcatcccgg agtatacgtt gaactcaacg ctgagctggc aggcacggga
     2221 agatttgtcg atgcaaacga ccttcacctg gtacggcaag cagcagccga agaagtacaa
     2281 ctataaaggt cagccagcgg ttggaccgga aaccaaagaa attagtcctt acagcattgt
     2341 tggcctgagc gcgacctggg atgtgacgaa gaatgtcagt ctgaccggcg gcgtggacaa
     2401 tctgttcgac aaacgtttgt ggcgtgcggg taatgcccag accacgggcg atttggcagg
     2461 ggccaactat atcgccggtg ccggggcgta tacctataac gagccgggac gtacgtggta
     2521 tatgagcgta aacacccact tctgatcatg gtacccgggg atcctctaga gtcgacctgc
     2581 aggcatgcaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
     2641 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
     2701 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
     2761 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
     2821 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     2881 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
     2941 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
     3001 tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct
     3061 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3121 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3181 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3241 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3301 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     3361 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3421 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3481 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3541 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3601 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3661 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3721 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3781 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3841 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3901 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3961 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc
     4021 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4081 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4141 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4201 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     4261 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4321 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     4381 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4441 attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat
     4501 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4561 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4621 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4681 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     4741 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4801 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4861 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4921 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4981 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5041 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5101 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5161 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5221 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5281 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5341 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5401 cgcatagggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5461 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5521 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gaaggagatg gcgcccaaca
     5581 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     5641 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     5701 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgc tcatgtttga
     5761 cagcttatca tcgatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc
     5821 ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg
     5881 gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg
     5941 cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg
     6001 gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc
     6061 tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac
     6121 ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga
     6181 tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg
     6241 ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc
     6301 gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc
     6361 ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta
     6421 agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga
     6481 gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc
     6541 ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga
     6601 ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg
     6661 gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg
     6721 ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagag
//
