LOCUS       Exported                5919 bp DNA     circular SYN 04-SEP-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5919)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5919)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             458..1252
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    1657..1678
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1693..1723
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1731..1747
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1755..1771
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1792..1810
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1840..1856
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1882..2601)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(2631..2659)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     primer_bind     complement(2661..2677)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2710..2728)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2738..2754)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3463..4125)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(4126..4897)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     promoter        5002..5053
                     /label=promoter for mob
     misc_feature    5020..5042
                     /label=RSA
                     /note="transfer origins (also called recombination site A 
                     [RSA]), is known as the specific site necessary for 
                     mobilization and recombination mediated by a Mob/Pre 
                     protein. "
ORIGIN
        1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
       61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
      121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
      181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
      241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
      301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
      361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
      421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
      481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta
     1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat
     1561 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1621 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     1681 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1741 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1801 tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacg gtatcgataa
     1861 gcttgatatc gaattcaaaa attacttgta cagctcgtcc atgccgagag tgatcccggc
     1921 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     1981 ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     2041 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     2101 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     2161 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     2221 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     2281 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     2341 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     2401 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     2461 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     2521 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     2581 ctcctcgccc ttgctcacca tgaatactgt ttcctgtgtg aaattccaca cattatacga
     2641 gccgatgatt aattgtcaag gatccactag ttctagagcg gccgccaccg cggtggagct
     2701 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     2761 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2821 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2881 tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2941 ttaaatcagc tcatttttta accaataggc cgtactgcga tgagtggcag ggcggggcgt
     3001 aattttttta aggcagttat tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg
     3061 ataataagcg gatgaatggc agaaattcga aagcaaattc gacccggtcg tcggttcagg
     3121 gcagggtcgt taaatagccg cttatgtcta ttgctggttt accggtttat tgactaccgg
     3181 aagcagtgtg accgtgtgct tctcaaatgc ctgaggccag tttgctcagg ctctccccgt
     3241 ggaggtaata attgacgata tgatcattta ttctgcctcc cagagcctga taaaaacggt
     3301 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     3361 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgaggcggct acagccgata
     3421 gtctggaaca gcgcacttac gggttgctgc gcaacccaag tgctaccggc gcggcagcgt
     3481 gacccgtgtc ggcggctcca acggctcgcc atcgtccaga aaacacggct catcgggcat
     3541 cggcaggcgc tgctgcccgc gccgttccca ttcctccgtt tcggtcaagg ctggcaggtc
     3601 tggttccatg cccggaatgc cgggctggct gggcggctcc tcgccggggc cggtcggtag
     3661 ttgctgctcg cccggataca gggtcgggat gcggcgcagg tcgccatgcc ccaacagcga
     3721 ttcgtcctgg tcgtcgtgat caaccaccac ggcggcactg aacaccgaca ggcgcaactg
     3781 gtcgcggggc tggccccacg ccacgcggtc attgaccacg taggccgaca cggtgccggg
     3841 gccgttgagc ttcacgacgg agatccagcg ctcggccacc aagtccttga ctgcgtattg
     3901 gaccgtccgc aaagaacgtc cgatgagctt ggaaagtgtc ttctggctga ccaccacggc
     3961 gttctggtgg cccatctgcg ccacgaggtg atgcagcagc attgccgccg tgggtttcct
     4021 cgcaataagc ccggcccacg cctcatgcgc tttgcgttcc gtttgcaccc agtgaccggg
     4081 cttgttcttg gcttgaatgc cgatttctct ggactgcgtg gccatgctta tctccatgcg
     4141 gtaggggtgc cgcacggttg cggcaccatg cgcaatcagc tgcaactttt cggcagcgcg
     4201 acaacaatta tgcgttgcgt aaaagtggca gtcaattaca gattttcttt aacctacgca
     4261 atgagctatt gcggggggtg ccgcaatgag ctgttgcgta cccccctttt ttaagttgtt
     4321 gatttttaag tctttcgcat ttcgccctat atctagttct ttggtgccca aagaagggca
     4381 cccctgcggg gttcccccac gccttcggcg cggctccccc tccggcaaaa agtggcccct
     4441 ccggggcttg ttgatcgact gcgcggcctt cggccttgcc caaggtggcg ctgccccctt
     4501 ggaacccccg cactcgccgc cgtgaggctc ggggggcagg cgggcgggct tcgcccttcg
     4561 actgccccca ctcgcatagg cttgggtcgt tccaggcgcg tcaaggccaa gccgctgcgc
     4621 ggtcgctgcg cgagccttga cccgccttcc acttggtgtc caaccggcaa gcgaagcgcg
     4681 caggccgcag gccggaggct tttccccaga gaaaattaaa aaaattgatg gggcaaggcc
     4741 gcaggccgcg cagttggagc cggtgggtat gtggtcgaag gctgggtagc cggtgggcaa
     4801 tccctgtggt caagctcgtg ggcaggcgca gcctgtccat cagcttgtcc agcagggttg
     4861 tccacgggcc gagcgaagcg agccagccgg tggccgctcg cggccatcgt ccacatatcc
     4921 acgggctggc aagggagcgc agcgaccgcg cagggcgaag cccggagagc aagcccgtag
     4981 ggcgccgcag ccgccgtagg cggtcacgac tttgcgaagc aaagtctagt gagtatactc
     5041 aagcattgag tggcccgccg gaggcaccgc cttgcgctgc ccccgtcgag ccggttggac
     5101 accaaaaggg aggggcaggc atggcggcat acgcgatcat gcgatgcaag aagctggcga
     5161 aaatgggcaa cgtggcggcc agtctcaagc acgcctaccg cgagcgcgag acgcccaacg
     5221 ctgacgccag caggacgcca gagaacgagc actgggcggc cagcagcacc gatgaagcga
     5281 tgggccgact gcgcgagttg ctgccagaga agcggcgcaa ggacgctgtg ttggcggtcg
     5341 agtacgtcat gacggccagc ccggaatggt ggaagtcggc cagccaagaa cagcaggcgg
     5401 cgttcttcga gaaggcgcac aagtggctgg cggacaagta cggggcggat cgcatcgtga
     5461 cggccagcat ccaccgtgac gaaaccagcc cgcacatgac cgcgttcgtg gtgccgctga
     5521 cgcaggacgg caggctgtcg gccaaggagt tcatcggcaa caaagcgcag atgacccgcg
     5581 accagaccac gtttgcggcc gctgtggccg atctagggct gcaacggggc atcgagggca
     5641 gcaaggcacg tcacacgcgc attcaggcgt tctacgaggc cctggagcgg ccaccagtgg
     5701 gccacgtcac catcagcccg caagcggtcg agccacgcgc ctatgcaccg cagggattgg
     5761 ccgaaaagct gggaatctca aagcgcgttg agacgccgga agccgtggcc gaccggctga
     5821 caaaagcggt tcggcagggg tatgagcctg ccctacaggc cgccgcagga gcgcgtgaga
     5881 tgcgcaagaa ggccgatcaa gcccaagaga cggcccgag
//
