LOCUS       Exported                6038 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 6038)
  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..6038
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
     CDS             1363..2076
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     1363..1382
                     /label=GFP_XbaI
     CDS             2077..2106
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     CDS             2116..2250
                     /codon_start=1
                     /label=TolAI
                     /translation="VSKATEQNDKLKRAIIISAVLHVILFAALIWSSFDENIEASAGGG
                     "
     primer_bind     2230..2271
                     /label=TonBII-Swap-F
     CDS             2251..2532
                     /codon_start=1
                     /label=TonBII
                     /translation="ISVTMVTPADLEPPQAVQPPPEPVVEPEPEPEPIPEPPKEAPVVI
                     EKPKPKPKPKPKPVKKVQEQPKRDVKPVESRPASPFENTAPARLTSSTA"
     CDS             2533..2838
                     /codon_start=1
                     /label=TonBIII
                     /translation="TAATSKPVTSVASGPRALSRNQPQYPARAQALRIEGQVKVKFDVT
                     PDGRVDNVQILSAKPANMFEREVKNAMRRWRYEPGKPGSGIVVNILFKINGTTEIQ"
     primer_bind     complement(2818..2859)
                     /label=TonBIII-R
     terminator      3065..3151
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3243..3270
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3289..3380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3381..4241
                     /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"
     rep_origin      4283..4738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      4849..5437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 tggtacccgg ggatcctcta gatttaagaa ggagatatac atatgagtaa aggagaagaa
     1381 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
     1441 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
     1501 atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac tttcgcgtat
     1561 ggtcttcaat gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt
     1621 gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac
     1681 aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa
     1741 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac
     1801 tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa
     1861 attagacaca acattgaaga tggaagcgtt caactagcag accattatca acaaaatact
     1921 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
     1981 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
     2041 gctgggatta cacatggcat ggatgaacta tacaaagcta gcatgactgg tggacagcaa
     2101 atgggtcgcg gatccgtgtc aaaggcaacc gaacaaaacg acaagctcaa gcgggcgata
     2161 attatttcag cagtgctgca tgtcatctta tttgcggcgc tgatctggag ttcgttcgat
     2221 gagaatatag aagcttcagc cggaggcggc atttctgtca cgatggttac gcctgctgat
     2281 ctcgaaccgc cacaagccgt tcagccgcca ccggagccgg tggtagagcc agaaccggaa
     2341 cctgagccga tccccgaacc gccaaaagaa gcaccggtgg tcattgaaaa gccgaagccg
     2401 aaacctaagc caaaaccgaa gccggtgaaa aaggtacagg agcagccaaa acgcgatgtc
     2461 aaacccgtag agtcgcgtcc ggcatcaccg tttgaaaata cggcaccggc acgcctgaca
     2521 tcaagtacag caacggctgc aaccagcaag ccggttacca gtgtggcttc aggaccacgc
     2581 gcattaagcc gtaatcagcc gcagtatccg gcacgagcac aggcattgcg cattgaaggg
     2641 caggttaaag ttaaatttga cgttacgccg gatggtcgcg tggataacgt acaaatcctc
     2701 tcagccaagc ctgcgaacat gtttgagcgt gaggtgaaaa atgcgatgcg cagatggcgt
     2761 tatgagccgg gtaagccagg cagtgggatt gtggtgaata tcctgtttaa aattaacggc
     2821 accaccgaaa ttcagtaagt cgacctgcag gcatgcaagc ttggctgttt tggcggatga
     2881 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     2941 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     3001 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     3061 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     3121 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     3181 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     3241 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
     3301 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3361 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3421 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3481 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3541 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3601 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     3661 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3721 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3781 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3841 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3901 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3961 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4021 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4081 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4141 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4201 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4261 catatatact ttagattgat ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4321 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4381 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4441 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4501 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4561 gagtccacgt tctttaatag tggactcttg ttccaaactt gaacaacact caaccctatc
     4621 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4681 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttaa
     4741 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4801 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4981 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5461 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5521 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5581 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     5641 ccttacgcat ctgtgcggta tttcacaccg catagggtca tggctgcgcc ccgacacccg
     5701 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5761 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5821 gcgaggcagc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     5881 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     5941 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     6001 tccggcgtag aggatctgct catgtttgac agcttatc
//
