LOCUS       pRL662-eGFP-SpeR        4718 bp DNA     circular SYN 27-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4718)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 30, 2024 from SnapGene Viewer 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      88..857
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             858..1520
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     2227..2243
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2253..2271
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2304..2320
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(2384..2431)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2496..3212)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     RBS             3220..3242
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(3274..3292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3344..3362)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3383..3399)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3407..3423
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(join(3431..3437,3438..3455,3456..3461))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3476..3497
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3694..4485)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(join(4674..4679,4680..4696,4697..4702))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 cgccctacgg gcttgctctc cgggcttcgc cctgcgcggt cgctgcgctc ccttgccagc
       61 ccgtggatat gtggacgatg gccgcgagcg gccaccggct ggctcgcttc gctcggcccg
      121 tggacaaccc tgctggacaa gctgatggac aggctgcgcc tgcccacgag cttgaccaca
      181 gggattgccc accggctacc cagccttcga ccacataccc accggctcca actgcgcggc
      241 ctgcggcctt gccccatcaa tttttttaat tttctctggg gaaaagcctc cggcctgcgg
      301 cctgcgcgct tcgcttgccg gttggacacc aagtggaagg cgggtcaagg ctcgcgcagc
      361 gaccgcgcag cggcttggcc ttgacgcgcc tggaacgacc caagcctatg cgagtggggg
      421 cagtcgaagg cgaagcccgc ccgcctgccc cccgagcctc acggcggcga gtgcgggggt
      481 tccaaggggg cagcgccacc ttgggcaagg ccgaaggccg cgcagtcgat caacaagccc
      541 cggaggggcc actttttgcc ggagggggag ccgcgccgaa ggcgtggggg aaccccgcag
      601 gggtgccctt ctttgggcac caaagaacta gatatagggc gaaatgcgaa agacttaaaa
      661 atcaacaact taaaaaaggg gggtacgcaa cagctcattg cggcaccccc cgcaatagct
      721 cattgcgtag gttaaagaaa atctgtaatt gactgccact tttacgcaac gcataattgt
      781 tgtcgcgctg ccgaaaagtt gcagctgatt gcgcatggtg ccgcaaccgt gcggcaccct
      841 accgcatgga gataagcatg gccacgcagt ccagagaaat cggcattcaa gccaagaaca
      901 agcccggtca ctgggtgcaa acggaacgca aagcgcatga ggcgtgggcc gggcttattg
      961 cgaggaaacc cacggcggca atgctgctgc atcacctcgt ggcgcagatg ggccaccaga
     1021 acgccgtggt ggtcagccag aagacacttt ccaagctcat cggacgttct ttgcggacgg
     1081 tccaatacgc agtcaaggac ttggtggccg agcgctggat ctccgtcgtg aagctcaacg
     1141 gccccggcac cgtgtcggcc tacgtggtca atgaccgcgt ggcgtggggc cagccccgcg
     1201 accagttgcg cctgtcggtg ttcagtgccg ccgtggtggt tgatcacgac gaccaggacg
     1261 aatcgctgtt ggggcatggc gacctgcgcc gcatcccgac cctgtatccg ggcgagcagc
     1321 aactaccgac cggccccggc gaggagccgc ccagccagcc cggcattccg ggcatggaac
     1381 cagacctgcc agccttgacc gaaacggagg aatgggaacg gcgcgggcag cagcgcctgc
     1441 cgatgcccga tgagccgtgt tttctggacg atggcgagcc gttggagccg ccgacacggg
     1501 tcacgctgcc gcgccggtag cacttgggtt gcgcagcaac ccgtaagtgc gctgttccag
     1561 actatcggct gtagccgcct cgccgcccta taccttgtct gcctccccgc gttgcgtcgc
     1621 ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc gctaacggat
     1681 tcaccgtttt tatcaggctc tgggaggcag aataaatgat catatcgtca attattacct
     1741 ccacggggag agcctgagca aactggcctc aggcatttga gaagcacacg gtcacactgc
     1801 ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     1861 gccctgaacc gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt
     1921 atcacttatt caggcgtagc accaggcgtt taagggcacc aataactgcc ttaaaaaaat
     1981 tacgccccgc cctgccactc atcgcagtcg gcctattggt taaaaaatga gctgatttaa
     2041 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca
     2101 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     2161 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     2221 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
     2281 agctccaccg cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattca
     2341 tcgatatcta gatctcgaat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga
     2401 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact
     2461 cagcttcctt tcgggctttg ttagcagccg gatcctttgt atagttcatc catgccatgt
     2521 gtaatcccag cagctgttac aaactcaaga aggaccatgt ggtctctctt ttcgttggga
     2581 tctttcgaaa gggcagattg tgtggacagg taatggttgt ctggtaaaag gacagggcca
     2641 tcgccaattg gagtattttg ttgataatgg tctgctagtt gaacgcttcc atcttcaatg
     2701 ttgtgtctaa ttttgaagtt agctttgatt ccattctttt gtttgtctgc catgatgtat
     2761 acattgtgtg agttatagtt gtattccaat ttgtgcccaa gaatgtttcc atcttcttta
     2821 aaatcaatac cttttaactc gattctatta acaagggtat caccttcaaa cttgacttca
     2881 gcacgtgtct tgtagttccc gtcatctttg aaaaatatag ttctttcctg tacataacct
     2941 tcgggcatgg cactcttgaa aaagtcatgc cgtttcatat gatccgggta tcttgaaaag
     3001 cactgaacac cataagtgaa agtagtgaca agtgttggcc atggaacagg tagttttcca
     3061 gtagtgcaaa taaatttaag ggtaagtttt ccgtatgttg catcaccttc accctctcca
     3121 ctgacagaaa atttgtgccc attaacatca ccatctaatt caacaagaat tgggacaact
     3181 ccagtgaaga gttcttctcc tttgctagcc atatgtatat ctccttctta aagttaaaca
     3241 aaattatttc tagagggaaa ccgttgtggt ctccctatag tgagtcgtat taatttcgcg
     3301 ggatcgagat ctcgaggggg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     3361 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3421 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3481 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3541 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3601 gcatggggta ccgagctcga attggccgcg gcgttgtgac aatttaccga acaactccgc
     3661 ggccgggaag ccgatctcgg cttgaacgaa ttgttatttg ccgactacct tggtgatctc
     3721 gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc
     3781 ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag
     3841 gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct
     3901 gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg
     3961 cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg
     4021 atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt
     4081 cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc
     4141 attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat
     4201 gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga
     4261 agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac
     4321 ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga
     4381 gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac
     4441 ctctgatagt tgagtcgata cttcggcgat caccgcttcc cccatcgttg ctgctccata
     4501 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
     4561 gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc
     4621 gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac
     4681 gaaccgaaca ggcttatgtc aattcgagct cggtaccc
//
