LOCUS       sequence-339586-        6784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FepA D34C . 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 6784)
  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 6784)
  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: pKP1583
FEATURES             Location/Qualifiers
     source          1..6784
                     /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(3642..3659)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3812..3898
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3990..4017
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4036..4127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4128..4196,4197..4988)
                     /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(4346..4365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5030..5485
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5117..5136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5327..5348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5596..6184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6085..6104
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6338..6355
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6521..6543
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6681..6700
                     /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 gccgaccgat actcctgttt cacatgacga tactattgtc gttaccgccg
     1441 ccgagcagaa cttacaggcg cctggcgttt cgaccatcac cgcatgcgaa atccgcaaaa
     1501 acccggttgc ccgcgatgtg tcgaagatca tccgtaccat gccaggcgtt aacctgaccg
     1561 gtaactccac cagtggtcag cgtgggaata accgacagat tgatattcgc ggtatgggtc
     1621 cggaaaacac gctgattttg attgacggca agccggtaag cagccgtaac tcggtgcgtc
     1681 agggctggcg tggcgagcgc gatacccgtg gtgatacttc ctgggtgcca cctgaaatga
     1741 ttgaacgtat tgaagttctg cgtggtccgg cagctgcgcg ttatggcaac ggcgcggcgg
     1801 gcggcgtggt taacatcatt accaaaaaag gcagcggcga gtggcacggc tcctgggacg
     1861 catatttcaa tgcgccagaa cataaagagg aaggtgccac caaacgcact aactttagcc
     1921 tgaccggtcc gctgggcgac gaattcagct tccgtttgta tggcaacctc gacaaaaccc
     1981 aggctgacgc gtgggatatc aaccagggcc atcagtccgc gcgtgccgga acgtatgcca
     2041 cgacgttacc agccgggcgc gaaggggtaa tcaacaaaga tattaatggc gtggtgcgct
     2101 gggatttcgc gccattgcaa tcgctggaac tggaagcagg ttacagccgc cagggtaacc
     2161 tgtatgcggg cgacacccag aataccaact ccgattccta tacccgctcg aaatatggcg
     2221 atgaaaccaa ccgtctgtat cgccagaact acgcgctgac ctggaacggt ggctgggata
     2281 acggcgtgac caccagcaac tgggtgcagt acgaacacac ccgtaactcg cgtattccgg
     2341 aaggtctggc gggcggtacc gaagggaaat ttaacgaaaa agcgacacag gatttcgtcg
     2401 atatcgatct tgatgacgtg atgctgcaca gcgaagttaa cctgccgatt gatttcctcg
     2461 ttaaccagac gctgacgctg ggtacggagt ggaatcagca acggatgaag gacttaagtt
     2521 ccaacaccca ggcactgacc ggaacgaata ccggtggcgc tattgatggc gtgagtacca
     2581 ccgaccgtag cccgtattca aaagcagaaa ttttctcgct gtttgccgaa aacaacatgg
     2641 agctgactga cagcaccatc gtaacgccgg ggctgcgttt cgatcatcac agtattgtcg
     2701 gcaataactg gagcccggcg ctgaacatat cgcaaggttt aggcgatgac ttcacgctga
     2761 aaatgggcat cgcccgtgct tataaagcgc cgagcctgta ccagactaac ccgaactaca
     2821 ttctctacag taaaggtcag ggttgctatg ccagcgcggg cggctgctat ctgcaaggta
     2881 acgatgacct gaaagcagaa accagcatca acaaagagat tggtctggag ttcaaacgcg
     2941 acgggtggct ggcgggcgtc acctggttcc gtaacgatta tcgcaataag attgaagcgg
     3001 gctatgtggc tgtagggcaa aacgcagtcg gcaccgatct ctatcagtgg gataacgtgc
     3061 cgaaagcggt ggttgaaggt ctggaaggat cgttaaacgt accggttagc gaaacggtga
     3121 tgtggaccaa taacatcact tatatgctga agagtgaaaa caaaaccacg ggcgaccgtt
     3181 tgtcgatcat cccggagtat acgttgaact caacgctgag ctggcaggca cgggaagatt
     3241 tgtcgatgca aacgaccttc acctggtacg gcaagcagca gccgaagaag tacaactata
     3301 aaggtcagcc agcggttgga ccggaaacca aagaaattag tccttacagc attgttggcc
     3361 tgagcgcgac ctgggatgtg acgaagaatg tcagtctgac cggcggcgtg gacaatctgt
     3421 tcgacaaacg tttgtggcgt gcgggtaatg cccagaccac gggcgatttg gcaggggcca
     3481 actatatcgc cggtgccggg gcgtatacct ataacgagcc gggacgtacg tggtatatga
     3541 gcgtaaacac ccacttctga tcatggtacc cggggatcct ctagagtcga cctgcaggca
     3601 tgcaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     3661 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     3721 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     3781 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     3841 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     3901 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     3961 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     4021 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4081 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4141 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4201 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4261 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4321 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     4381 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4441 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4501 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4561 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4621 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4681 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4741 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4801 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4861 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4921 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4981 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta cgcgccctgt
     5041 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5101 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5161 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5221 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     5281 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5341 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg
     5401 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5461 aacaaaatat taacgtttac aatttaaaag gatctaggtg aagatccttt ttgataatct
     5521 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5581 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5641 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5701 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     5761 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5821 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5881 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5941 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6001 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6061 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6121 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6181 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6241 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6301 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6361 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     6421 agggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     6481 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     6541 aggttttcac cgtcatcacc gaaacgcgcg aggcagaagg agatggcgcc caacagtccc
     6601 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     6661 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     6721 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg tttgacagct
     6781 tatc
//
