LOCUS       sequence-339798-        6769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FepA G54C, 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: pKP1627
FEATURES             Location/Qualifiers
     source          1..6769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3627..3644)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3797..3883
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3975..4002
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4021..4112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4113..4181,4182..4973)
                     /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(4331..4350)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5015..5470
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5102..5121)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5312..5333
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5581..6169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6070..6089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6323..6340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6506..6528
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6666..6685
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattcacca
     1321 tgaacaagaa gattcattcc ctggccttgt tggtcaatct ggggatttat ggggtagcgc
     1381 aggcacaaga gccgacggat actcctgttt cacatgacac cgccgccgag cagaacttac
     1441 aggcgcctgg cgtttcgacc atcaccgcag atgaaatccg caaaaacccg gttgcccgcg
     1501 atgtgtcgaa gatcatccgt accatgccat gcgttaacct gaccggtaac tccaccagtg
     1561 gtcagcgtgg gaataaccga cagattgata ttcgcggtat gggtccggaa aacacgctga
     1621 ttttgattga cggcaagccg gtaagcagcc gtaactcggt gcgtcagggc tggcgtggcg
     1681 agcgcgatac ccgtggtgat acttcctggg tgccacctga aatgattgaa cgtattgaag
     1741 ttctgcgtgg tccggcagct gcgcgttatg gcaacggcgc ggcgggcggc gtggttaaca
     1801 tcattaccaa aaaaggcagc ggcgagtggc acggctcctg ggacgcatat ttcaatgcgc
     1861 cagaacataa agaggaaggt gccaccaaac gcactaactt tagcctgacc ggtccgctgg
     1921 gcgacgaatt cagcttccgt ttgtatggca acctcgacaa aacccaggct gacgcgtggg
     1981 atatcaacca gggccatcag tccgcgcgtg ccggaacgta tgccacgacg ttaccagccg
     2041 ggcgcgaagg ggtaatcaac aaagatatta atggcgtggt gcgctgggat ttcgcgccat
     2101 tgcaatcgct ggaactggaa gcaggttaca gccgccaggg taacctgtat gcgggcgaca
     2161 cccagaatac caactccgat tcctataccc gctcgaaata tggcgatgaa accaaccgtc
     2221 tgtatcgcca gaactacgcg ctgacctgga acggtggctg ggataacggc gtgaccacca
     2281 gcaactgggt gcagtacgaa cacacccgta actcgcgtat tccggaaggt ctggcgggcg
     2341 gtaccgaagg gaaatttaac gaaaaagcga cacaggattt cgtcgatatc gatcttgatg
     2401 acgtgatgct gcacagcgaa gttaacctgc cgattgattt cctcgttaac cagacgctga
     2461 cgctgggtac ggagtggaat cagcaacgga tgaaggactt aagttccaac acccaggcac
     2521 tgaccggaac gaataccggt ggcgctattg atggcgtgag taccaccgac cgtagcccgt
     2581 attcaaaagc agaaattttc tcgctgtttg ccgaaaacaa catggagctg actgacagca
     2641 ccatcgtaac gccggggctg cgtttcgatc atcacagtat tgtcggcaat aactggagcc
     2701 cggcgctgaa catatcgcaa ggtttaggcg atgacttcac gctgaaaatg ggcatcgccc
     2761 gtgcttataa agcgccgagc ctgtaccaga ctaacccgaa ctacattctc tacagtaaag
     2821 gtcagggttg ctatgccagc gcgggcggct gctatctgca aggtaacgat gacctgaaag
     2881 cagaaaccag catcaacaaa gagattggtc tggagttcaa acgcgacggg tggctggcgg
     2941 gcgtcacctg gttccgtaac gattatcgca ataagattga agcgggctat gtggctgtag
     3001 ggcaaaacgc agtcggcacc gatctctatc agtgggataa cgtgccgaaa gcggtggttg
     3061 aaggtctgga aggatcgtta aacgtaccgg ttagcgaaac ggtgatgtgg accaataaca
     3121 tcacttatat gctgaagagt gaaaacaaaa ccacgggcga ccgtttgtcg atcatcccgg
     3181 agtatacgtt gaactcaacg ctgagctggc aggcacggga agatttgtcg atgcaaacga
     3241 ccttcacctg gtacggcaag cagcagccga agaagtacaa ctataaaggt cagccagcgg
     3301 ttggaccgga aaccaaagaa attagtcctt acagcattgt tggcctgagc gcgacctggg
     3361 atgtgacgaa gaatgtcagt ctgaccggcg gcgtggacaa tctgttcgac aaacgtttgt
     3421 ggcgtgcggg taatgcccag accacgggcg atttggcagg ggccaactat atcgccggtg
     3481 ccggggcgta tacctataac gagccgggac gtacgtggta tatgagcgta aacacccact
     3541 tctgatcatg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttggctgt
     3601 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     3661 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     3721 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     3781 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     3841 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     3901 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     3961 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     4021 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4081 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4141 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4201 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4261 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4321 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     4381 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4441 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4501 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4561 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4621 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4681 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4741 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4801 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4861 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4921 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4981 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc
     5041 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5101 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5161 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5221 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5281 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5341 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5401 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5461 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5701 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5821 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5881 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5941 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6001 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6061 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6121 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6181 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6241 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6301 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     6361 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt catggctgcg
     6421 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     6481 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     6541 tcaccgaaac gcgcgaggca gaaggagatg gcgcccaaca gtcccccggc cacggggcct
     6601 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     6661 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     6721 gccacgatgc gtccggcgta gaggatctgc tcatgtttga cagcttatc
//
