LOCUS       Exported                6601 bp ds-DNA     circular SYN 27-APR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6601)
  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..6601
                     /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     2964..2983
                     /label=pBAD Forward
     primer_bind     3053..3092
                     /label=GFP-TonB_F1
     gene            3073..3780
                     /label=msGFP
     primer_bind     3073..3092
                     /label=GFP-TonB-XbaI
     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"
     misc_feature    3796..3891
                     /label=TonB domain I
     misc_feature    3892..3981
                     /label=TonB linker between domain I and II
     primer_bind     complement(3962..4001)
                     /label=Reverse Primer for TonB
     primer_bind     3962..4001
                     /label=Forward primer for TolA
     misc_feature    3982..4770
                     /label=TolA domain II
     primer_bind     4751..4790
                     /label=Forward Primer for TonB
     primer_bind     complement(4751..4790)
                     /label=Reverse Primer for TolA
     misc_feature    4771..4849
                     /label=TonB Linker between domain II and III
     primer_bind     4850..4889
                     /label=TonBII-TolAIII(100)-F
     gene            4850..4869
                     /label=tonB
     gene            4870..5175
                     /label=TolAIII
     primer_bind     complement(5156..5191)
                     /label=TolAIII-R
     terminator      5384..5470
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5562..5589
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5608..5699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5700..6560
                     /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(6369..6382)
                     /label=TolAII-TolAIII-F
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 gttctatgac ccttgattta cctcgccgct tcccctggcc gacgttactt
     3841 tcggtctgca ttcatggtgc tgttgtggcg ggtctgctct atacctcggt acatcaggtt
     3901 attgaactac ctgcgcctgc gcagccgatt tctgtcacga tggttacgcc tgctgatctc
     3961 gaaccgccac aagccgttca gtcgtccatc gacgctgtca tggttgattc aggtgcggta
     4021 gttgagcagt acaaacgcat gcaaagccag gaatcaagcg cgaagcgttc tgatgaacag
     4081 cgcaagatga aggaacagca ggctgctgaa gaactgcgtg agaaacaagc ggctgaacag
     4141 gaacgcctga agcaacttga gaaagagcgg ttagcggctc aggagcagaa aaagcaggct
     4201 gaagaagccg caaaacaggc cgagttaaag cagaagcaag ctgaagaggc ggcagcgaaa
     4261 gcggcggcag atgctaaagc gaaggccgaa gcagatgcta aagctgcgga agaagcagcg
     4321 aagaaagcgg ctgcagacgc aaagaaaaaa gcagaagcag aagccgccaa agccgcagcc
     4381 gaagcgcaga aaaaagccga ggcagccgct gcggcactga agaagaaagc ggaagcggca
     4441 gaagcagctg cagctgaagc aagaaagaaa gcggcaactg aagctgctga aaaagccaaa
     4501 gcagaagctg agaagaaagc ggctgctgaa aaggctgcag ctgataagaa agcggcagca
     4561 gagaaagctg cagccgacaa aaaagcagca gaaaaagcgg ctgctgaaaa ggcagcagct
     4621 gataagaaag cagcggcaga aaaagccgcc gcagacaaaa aagcggcagc ggcaaaagct
     4681 gcagctgaaa aagccgctgc agcaaaagcg gccgcagagg cagatgatat tttcggtgag
     4741 ctaagctctg gtaagaatgc accgaaaacg gtacaggagc agccaaaacg cgatgtcaaa
     4801 cccgtagagt cgcgtccggc atcaccgttt gaaaatacgg caccggcacg cctgacatca
     4861 agtacagcac ctgccgggag tggtaatact aaaaacaatg gcgcatcagg ggccgatatc
     4921 aataactatg ccgggcagat taaatctgct atcgaaagta agttctatga cgcatcgtcc
     4981 tatgcaggca aaacctgtac gctgcgcata aaactggcac ccgatggtat gttactggat
     5041 atcaaacctg aaggtggcga tcccgcactt tgtcaggctg cgttggcagc agctaaactt
     5101 gcgaagatcc cgaaaccacc aagccaggca gtatatgaag tgttcaaaaa cgcgccattg
     5161 gacttcaaac cgtaaaagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     5221 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     5281 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     5341 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     5401 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     5461 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     5521 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     5581 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     5641 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5701 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5761 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5821 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5881 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5941 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6001 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6061 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6121 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6181 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6241 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6301 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6361 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6421 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6481 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6541 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6601 t
//
