LOCUS       Exported                5809 bp ds-DNA     circular SYN 08-DEC-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5809)
  AUTHORS   bioc1520
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Aug 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5809
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      567..1155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1852..2730)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2757..3041
                     /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     3053..3092
                     /label=GFP-TonB_F1
     gene            3073..3780
                     /label=msGFP
     primer_bind     3761..3815
                     /label=GFP-TonB_F2
     primer_bind     complement(3761..3815)
                     /label=GFP-TonB_R1
     misc_feature    3781..3795
                     /label=Linker
     CDS             3781..3795
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="GGGGS"
     CDS             3796..3888
                     /codon_start=1
                     /label=TonB - Domain I
                     /translation="TLDLPRRFPWPTLLSVCIHGAVVAGLLYTSV"
     misc_feature    3889..3978
                     /label=Linker between domain I and II
     primer_bind     3959..3998
                     /label=Forward Primer (TonB without II domain)
     primer_bind     complement(3959..3998)
                     /label=Reverse Primer (TonB without domain II)
     misc_feature    3979..4167
                     /label=Linker between domain II and domain III
     misc_feature    4168..4383
                     /label=Domain III
     primer_bind     complement(4364..4403)
                     /label=GFP-TonB_R2
     terminator      4592..4678
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4770..4797
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4816..4907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4908..5768
                     /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"
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 ttgatttggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaacttga acaacactca accctatctc gggctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgttta caatttaaaa ggatctaggt gaagatcctt
      481 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
      541 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
      601 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      661 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
      721 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
      781 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      841 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      901 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      961 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1021 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1081 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1141 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     1201 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     1261 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     1321 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     1381 tcacaccgca tagggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     1441 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     1501 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcaa ggagatggcg
     1561 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     1621 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     1681 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctgctca
     1741 tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag cagggattct
     1801 gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa
     1861 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
     1921 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
     1981 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     2041 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag
     2101 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     2161 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     2221 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     2281 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     2341 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     2401 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     2461 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     2521 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     2581 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     2641 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     2701 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga
     2761 aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc
     2821 gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc
     2881 catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta
     2941 tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc
     3001 tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt tttgggctag
     3061 caggaggaat tcatggatag cactgagagc ctgttcaccg gggtggtgcc catcctggtc
     3121 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg cgagggcgat
     3181 gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3241 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     3301 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3361 accatcacct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa gttcgagggc
     3421 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3481 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     3541 cagaagaacg gcatcaaggc caacttcaag atccgccaca acgtggagga cggcagcgtg
     3601 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3661 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     3721 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctggaggcag tggcggttct
     3781 ggtggtggtg gttctaccct tgatttacct cgccgcttcc cctggccgac gttactttcg
     3841 gtctgcattc atggtgctgt tgtggcgggt ctgctctata cctcggtaca tcaggttatt
     3901 gaactacctg cgcctgcgca gccgatttct gtcacgatgg ttacgcctgc tgatctcgaa
     3961 ccgccacaag ccgttcaggt acaggagcag ccaaaacgcg atgtcaaacc cgtagagtcg
     4021 cgtccggcat caccgtttga aaatacggca ccggcacgcc tgacatcaag tacagcaacg
     4081 gctgcaacca gcaagccggt taccagtgtg gcttcaggac cacgcgcatt aagccgtaat
     4141 cagccgcagt atccggcacg agcacaggca ttgcgcattg aagggcaggt taaagttaaa
     4201 tttgacgtta cgccggatgg tcgcgtggat aacgtacaaa tcctctcagc caagcctgcg
     4261 aacatgtttg agcgtgaggt gaaaaatgcg atgcgcagat ggcgttatga gccgggtaag
     4321 ccaggcagtg ggattgtggt gaatatcctg tttaaaatta acggcaccac cgaaattcag
     4381 taaaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     4441 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     4501 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     4561 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     4621 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     4681 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     4741 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     4801 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4861 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4921 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4981 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5041 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5101 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     5161 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5221 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5281 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5341 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5401 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5461 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5521 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5581 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5641 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5701 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5761 attggtaact gtcagaccaa gtttactcat atatacttta gattgattt
//
