LOCUS       Exported                3523 bp DNA     circular SYN 14-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3523)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2023 from SnapGene 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..861
                     /codon_start=1
                     /label=mVenus full ORF
                     /translation="MRGSHHHHHHGMASMVSKGEELFTGVVPILVELDGDVNGHKFSVS
                     GEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMP
                     EGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSH
                     NVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKL
                     SKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     misc_feature    100..102
                     /label=start
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..141
                     /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="MAS"
     CDS             145..859
                     /codon_start=1
                     /label=mVenus 206k george
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     terminator      938..985
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             1012..1080
                     /label=LacZ alpha
     rep_origin      1082..1537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      1099..1405
                     /label=F1 ori
     promoter        1563..1667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1668..2528
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             1866..2525
                     /label=AmpR
     rep_origin      2623..3305
                     /label=ColE1 origin
     rep_origin      2699..3287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catggtgagc aagggcgagg agctgttcac cggggtggtg
      181 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      241 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
      301 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc
      361 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      421 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      481 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      541 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
      601 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      661 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      721 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
      781 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
      841 atggacgagc tgtacaagta agaattcgaa gcttgatccg gctgctaaca aagcccgaaa
      901 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
      961 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat ctggcgtaat
     1021 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1081 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1141 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1201 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1261 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1321 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1381 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1441 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1501 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     1561 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1681 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1741 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1801 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1861 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1921 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1981 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2041 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2101 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2161 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2221 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2281 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2341 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2401 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2461 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2521 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2581 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2641 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2701 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2761 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2821 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2881 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2941 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3001 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3061 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3121 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3181 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3241 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3301 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3361 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3421 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     3481 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cag
//
