LOCUS       sequence-337287-        6784 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express FepA D12C . 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 Apr 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: pKP1410
FEATURES             Location/Qualifiers
     source          1..6784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2323..2340)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2493..2579
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2671..2698
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2717..2808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2809..2877,2878..3669)
                     /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(3027..3046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3711..4166
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3798..3817)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4008..4029
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4277..4865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4766..4785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5019..5036
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5202..5224
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5362..5381
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(5561..6439)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        6466..6750
                     /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     6673..6692
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 atgaacaaga agattcattc cctggccttg ttggtcaatc tggggattta tggggtagcg
       61 caggcacaag agccgaccga tactcctgtt tcacatgact gtactattgt cgttaccgcc
      121 gccgagcaga acttacaggc gcctggcgtt tcgaccatca ccgcagatga aatccgcaaa
      181 aacccggttg cccgcgatgt gtcgaagatc atccgtacca tgccaggcgt taacctgacc
      241 ggtaactcca ccagtggtca gcgtgggaat aaccgacaga ttgatattcg cggtatgggt
      301 ccggaaaaca cgctgatttt gattgacggc aagccggtaa gcagccgtaa ctcggtgcgt
      361 cagggctggc gtggcgagcg cgatacccgt ggtgatactt cctgggtgcc acctgaaatg
      421 attgaacgta ttgaagttct gcgtggtccg gcagctgcgc gttatggcaa cggcgcggcg
      481 ggcggcgtgg ttaacatcat taccaaaaaa ggcagcggcg agtggcacgg ctcctgggac
      541 gcatatttca atgcgccaga acataaagag gaaggtgcca ccaaacgcac taactttagc
      601 ctgaccggtc cgctgggcga cgaattcagc ttccgtttgt atggcaacct cgacaaaacc
      661 caggctgacg cgtgggatat caaccagggc catcagtccg cgcgtgccgg aacgtatgcc
      721 acgacgttac cagccgggcg cgaaggggta atcaacaaag atattaatgg cgtggtgcgc
      781 tgggatttcg cgccattgca atcgctggaa ctggaagcag gttacagccg ccagggtaac
      841 ctgtatgcgg gcgacaccca gaataccaac tccgattcct atacccgctc gaaatatggc
      901 gatgaaacca accgtctgta tcgccagaac tacgcgctga cctggaacgg tggctgggat
      961 aacggcgtga ccaccagcaa ctgggtgcag tacgaacaca cccgtaactc gcgtattccg
     1021 gaaggtctgg cgggcggtac cgaagggaaa tttaacgaaa aagcgacaca ggatttcgtc
     1081 gatatcgatc ttgatgacgt gatgctgcac agcgaagtta acctgccgat tgatttcctc
     1141 gttaaccaga cgctgacgct gggtacggag tggaatcagc aacggatgaa ggacttaagt
     1201 tccaacaccc aggcactgac cggaacgaat accggtggcg ctattgatgg cgtgagtacc
     1261 accgaccgta gcccgtattc aaaagcagaa attttctcgc tgtttgccga aaacaacatg
     1321 gagctgactg acagcaccat cgtaacgccg gggctgcgtt tcgatcatca cagtattgtc
     1381 ggcaataact ggagcccggc gctgaacata tcgcaaggtt taggcgatga cttcacgctg
     1441 aaaatgggca tcgcccgtgc ttataaagcg ccgagcctgt accagactaa cccgaactac
     1501 attctctaca gtaaaggtca gggttgctat gccagcgcgg gcggctgcta tctgcaaggt
     1561 aacgatgacc tgaaagcaga aaccagcatc aacaaagaga ttggtctgga gttcaaacgc
     1621 gacgggtggc tggcgggcgt cacctggttc cgtaacgatt atcgcaataa gattgaagcg
     1681 ggctatgtgg ctgtagggca aaacgcagtc ggcaccgatc tctatcagtg ggataacgtg
     1741 ccgaaagcgg tggttgaagg tctggaagga tcgttaaacg taccggttag cgaaacggtg
     1801 atgtggacca ataacatcac ttatatgctg aagagtgaaa acaaaaccac gggcgaccgt
     1861 ttgtcgatca tcccggagta tacgttgaac tcaacgctga gctggcaggc acgggaagat
     1921 ttgtcgatgc aaacgacctt cacctggtac ggcaagcagc agccgaagaa gtacaactat
     1981 aaaggtcagc cagcggttgg accggaaacc aaagaaatta gtccttacag cattgttggc
     2041 ctgagcgcga cctgggatgt gacgaagaat gtcagtctga ccggcggcgt ggacaatctg
     2101 ttcgacaaac gtttgtggcg tgcgggtaat gcccagacca cgggcgattt ggcaggggcc
     2161 aactatatcg ccggtgccgg ggcgtatacc tataacgagc cgggacgtac gtggtatatg
     2221 agcgtaaaca cccacttctg atcatggtac ccggggatcc tctagagtcg acctgcaggc
     2281 atgcaagctt ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc
     2341 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
     2401 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
     2461 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     2521 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     2581 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
     2641 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
     2701 cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2761 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2821 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2881 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2941 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3001 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     3061 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3121 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3181 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3241 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3301 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3361 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3421 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3481 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3541 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3601 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3661 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt acgcgccctg
     3721 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3781 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3841 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3901 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3961 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4021 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     4081 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4141 taacaaaata ttaacgttta caatttaaaa ggatctaggt gaagatcctt tttgataatc
     4201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4981 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5041 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5101 tagggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5161 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5221 gaggttttca ccgtcatcac cgaaacgcgc gaggcagaag gagatggcgc ccaacagtcc
     5281 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     5341 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     5401 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc
     5461 ttatcatcga tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     5521 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     5581 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     5641 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     5701 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     5761 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     5821 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     5881 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     5941 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     6001 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     6061 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     6121 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     6181 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     6241 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     6301 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     6361 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     6421 cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc
     6481 catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac
     6541 cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa
     6601 cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg
     6661 tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct acctgacgct
     6721 ttttatcgca actctctact gtttctccat acccgttttt ttgggctagc aggaggaatt
     6781 cacc
//
