LOCUS       pAC17V5a        8100 bp DNA     circular SYN 14-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   urn.local...4u9-8o9e0sl
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8100)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 24, 2023 from SnapGene 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             255..848
     CDS             1596..2252
     oriT            2204..2291
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      2633..3027
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     primer_bind     3405..3422
                     /label=AC_06
     misc_feature    3496..3638
                     /label=conS
     primer_bind     3515..3534
                     /label=AC_75
     primer_bind     3541..3560
                     /label=AC_09
     primer_bind     complement(3557..3576)
                     /label=AC_69
     primer_bind     complement(3573..3592)
                     /label=AC_019
     primer_bind     3577..3598
                     /label=AC_71
     misc_feature    3643..3724
     promoter        3725..3783
                     /label=CP25
     primer_bind     3725..3740
                     /label=AC_74
     protein_bind    3784..3808
                     /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)."
     protein_bind    3784..3808
     protein_bind    3784..3808
                     /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)."
     RBS             3819..3826
     CDS             3835..4551
                     /codon_start=1
                     /label=GFP optim1
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     3933..3952
                     /label=AC_022
     primer_bind     complement(4019..4038)
                     /label=AC_023
     terminator      4556..4638
     misc_feature    4650..4674
     misc_feature    complement(4675..4694)
     misc_feature    4695..4714
     primer_bind     complement(4710..4729)
                     /label=AC_114
     misc_feature    complement(4715..4729)
     primer_bind     4730..4749
                     /label=AC_70
     misc_feature    complement(4735..5745)
     CDS             complement(4735..5745)
                     /codon_start=1
                     /label=SpecR
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(4735..5526)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    complement(4745..4764)
     primer_bind     complement(4746..4765)
                     /label=AC_07
     misc_feature    5631..5650
     promoter        complement(5746..5879)
     rep_origin      5880
     rep_origin      complement(5881..5905)
                     /direction=LEFT
     misc_feature    complement(5881..5905)
     primer_bind     5881..5903
                     /label=AC_018
     primer_bind     complement(5881..5903)
                     /label=AC_010
     rep_origin      join(5906..8100,1..3463)
                     /label=RSF1010
     CDS             complement(5906..6856)
     CDS             complement(5906..6757)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRKRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     primer_bind     complement(5928..5946)
                     /label=AC_05
     CDS             complement(6744..7583)
     CDS             complement(6744..7583)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             6819..7586
     primer_bind     7575..7594
                     /label=AC_59
     primer_bind     complement(7575..7594)
                     /label=AC_60
     CDS             complement(join(7994..8100,1..538))
     CDS             complement(join(8094..8100,1..2123))
     CDS             complement(join(8094..8100,1..965))
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDAPTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGHDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
ORIGIN
        1 ctgaaatctg gcccgcccct gtccatgcct cgctggcggg gtgccggtgc ccgtgccagc
       61 tcggcccgcg caagctggac gctgggcaga cccatgacct tgctgacggt gcgctcgatg
      121 taatccgctt cgtggccggg cttgcgctct gccagcgctg ggctggcctc ggccatggcc
      181 ttgccgattt cctcggcact gcggccccgg ctggccagct tctgcgcggc gataaagtcg
      241 cacttgctga ggtcatgacc gaagcgcttg accagcccgg ccatctcgct gcggtactcg
      301 tccagcgccg tgcgccggtg gcggctaagc tgccgctcgg gcagttcgag gctggccagc
      361 ctgcgggcct tctcctgctg ccgctgggcc tgctcgatct gctggccagc ctgctgcacc
      421 agcgccgggc cagcggtggc ggtcttgccc ttggattcac gcagcagcac ccacggctga
      481 taaccggcgc gggtggtgtg cttgtccttg cggttggtga agcccgccaa gcggccatag
      541 tggcggctgt cggcgctggc cgggtcggcg tcgtactcgc tggccagcgt ccgggcaatc
      601 tgcccccgaa gttcaccgcc tgcggcgtcg gccaccttga cccatgcctg atagttcttc
      661 gggctggttt ccactaccag ggcaggctcc cggccctcgg ctttcatgtc atccaggtca
      721 aactcgctga ggtcgtccac cagcaccaga ccatgccgct cctgctcggc gggcctgata
      781 tacacgtcat tgccctgggc attcatccgc ttgagccatg gcgtgttctg gagcacttcg
      841 gcggctgacc attcccggtt catcatctgg ccggtgggtg cgtccctgac gccgatatcg
      901 aagcgctcac agcccatggc cttgagctgt cggcctatgg cctgcaaagt cctgtcgttc
      961 ttcatcgggc caccaagcgc agccagatcg agccgtcctc ggttgtcagt ggcgtcaggt
     1021 cgagcaagag caacgatgcg atcagcagca ccaccgtagg catcatggaa gccagcatca
     1081 cggttagcca tagcttccag tgccaccccc gcgacgcgct ccgggcgctc tgcgcggcgc
     1141 tgctcacctc ggcggctacc tcccgcaact ctttggccag ctccacccat gccgcccctg
     1201 tctggcgctg ggctttcagc cactccgccg cctgcgcctc gctggcctgc ttggtctggc
     1261 tcatgacctg ccgggcttcg tcggccagtg tcgccatgct ctgggccagc ggttcgatct
     1321 gctccgctaa ctcgttgatg cctctggatt tcttcactct gtcgattgcg ttcatggtct
     1381 attgcctccc ggtattcctg taagtcgatg atctgggcgt tggcggtgtc gatgttcagg
     1441 gccacgtctg cccggtcggt gcggatgccc cggccttcca tctccaccac gttcggcccc
     1501 aggtgaacac cgggcaggcg ctcgatgccc tgcgcctcaa gtgttctgtg gtcaatgcgg
     1561 gcgtcgtggc cagcccgctc taatgcccgg ttggcatggt cggcccatgc ctcgcgggtc
     1621 tgctcaagcc atgccttggg cttgagcgct tcggtcttct gtgccccgcc cttctccggg
     1681 gtcttgccgt tgtaccgctt gaaccactga gcggcgggcc gctcgatgcc gtcattgatc
     1741 cgctcggaga tcatcaggtg gcagtgcggg ttctcgccgc caccggcatg gatggccagc
     1801 gtatacggca ggcgctcggc accggtcagg tgctgggcga actcggacgc cagcgccttc
     1861 tgctggtcga gggtcagctc gaccggcagg gcaaattcga cctccttgaa cagccgccca
     1921 ttggcgcgtt catacaggtc ggcagcatcc cagtagtcgg cgggccgctc gacgaactcc
     1981 ggcatgtgcc cggattcggc gtgcaagact tcatccatgt cgcgggcata cttgccttcg
     2041 cgctggatgt agtcggcctt ggccctggcc gattggccgc ccgacctgct gccggttttc
     2101 gccgtaaggt gataaatcgc catgctgcct cgctgttgct tttgcttttc ggctccatgc
     2161 aatggccctc ggagagcgca ccgcccgaag ggtggccgtt aggccagttt ctcgaagaga
     2221 aaccggtaag tgcgccctcc cctacaaagt agggtcggga ttgccgccgc tgtgcctcca
     2281 tgatagccta cgagacagca cattaacaat ggggtgtcaa gatggttaag gggagcaaca
     2341 aggcggcgga tcggctggcc aagctcgaag aacaacgagc gcgaatcaat gccgaaattc
     2401 agcgggagcg ggcaagggaa cagcagcaag agcgcaagaa cgaaacaagg cgcaaggtgc
     2461 tggtgggggc catgattttg gccaaggtga acagcagcga gtggccggag gatcggctca
     2521 tggcggcaat ggatgcgtac cttgaacgcg accacgaccg cgccttgttc ggtctgccgc
     2581 cacgccagaa ggatgagccg ggctgaatga tcgaccgaga caggccctgc ggggctgcac
     2641 acgcgccccc acccttcggg tagggggaaa ggccgctaaa gcggctaaaa gcgctccagc
     2701 gtatttctgc ggggtttggt gtggggttta gcgggctttg cccgcctttc cccctgccgc
     2761 gcagcggtgg ggcggtgtgt agcctagcgc agcgaataga ccagctatcc ggcctctggc
     2821 cgggcatatt gggcaagggc agcagcgccc cacaagggcg ctgataaccg cgcctagtgg
     2881 attattctta gataatcatg gatggatttt tccaacaccc cgccagcccc cgcccctgct
     2941 gggtttgcag gtttgggggc gtgacagtta ttgcaggggt tcgtgacagt tattgcaggg
     3001 gggcgtgaca gttattgcag gggttcgtga cagttagtac gggagtgacg ggcactggct
     3061 ggcaatgtct agcaacggca ggcatttcgg ctgagggtaa aagaactttc cgctaagcga
     3121 tagactgtat gtaaacacag tattgcaagg acgcggaaca tgcctcatgt ggcggccagg
     3181 acggccagcc gggatcggga tactggtcgt taccagagcc accgacccga gcaaaccctt
     3241 ctctatcaga tcgttgacga gtattacccg gcattcgctg cgcttatggc agagcaggga
     3301 aaggaattgc cgggctatgt gcaacgggaa tttgaagaat ttctccaatg cgggcggctg
     3361 gagcatggct ttctacgggt tcgctgcgag tcttgccacg ccgagcacct ggtcgctttc
     3421 agaaatcaat ctaaagtata tatgagtaaa cttggtctga caggcccctg aattcgcatc
     3481 tagactgatg agacgtggta gagccacaaa cagccggtac aagcaacgat ctccaggacc
     3541 atctgaatca tgcgcggatg acacgaactc acgacggcga tcacagacat taacccacag
     3601 tacagacact gcgacaacgt ggcaattcgt cgcaatacaa cgtcggtcag tttcacctga
     3661 tttacgtaaa aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac
     3721 tgtcctttgg cagtttattc ttgacatgta gtgagggggc tggtataatc acatagtact
     3781 gttggaattg tgagcggata acaattccat acagaaacag aggagatatt acatatgagt
     3841 aaaggagaag agcttttcac aggagttgtc ccaatcctcg tggaattaga cggtgatgtt
     3901 aatgggcaca agttctctgt cagtggagag ggtgaaggcg acgcaacata tggcaagctg
     3961 acccttaaat ttatttgcac cacgggtaaa ctacctgttc catggccaac actggtcact
     4021 acgttcgggt atggggttca gtgctttgcg cgctacccag atcacatgaa acagcacgac
     4081 tttttcaaga gtgcaatgcc cgaaggctat gtacaggaga gaaccatctt ttttaaggat
     4141 gacggcaact ataagacacg cgccgaagtg aagttcgagg gtgataccct tgttaataga
     4201 atcgagttaa agggtattga ctttaaggaa gatggaaata ttttaggcca caaactggaa
     4261 tataactata actcccataa tgtgtacatt atggccgaca agcaaaagaa cggtatcaag
     4321 gttaacttca agatcagaca caacattgag gatggaagcg ttcaactagc cgaccattac
     4381 caacaaaaca ccccaattgg cgatgggcct gtgctgttac cagacaacca ttacctgtcc
     4441 actcaatctg ccctttcgaa agatcccaac gaaaagcgcg accacatggt ccttcttgag
     4501 tttgtcacgg ctgctgggat tacacacggc atggatgaac tatacaaata aatccgtaat
     4561 cgttaatccg caaataacgt aaaaacccgc ttcggcgggt ttttttatgg ggggagttta
     4621 gggaaagagc atttgtcagc tgcgtctcaa gcagttacag agatgttacg aacccccagg
     4681 acatccgaga atgcgaggcg atggagggta caaccgggag agtgttcacg ccgcttattt
     4741 gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc
     4801 gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac
     4861 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     4921 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     4981 atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     5041 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     5101 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     5161 gaagataccc gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     5221 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     5281 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     5341 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt
     5401 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     5461 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     5521 cctcataatg tttaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     5581 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
     5641 agactgtacc ccaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc
     5701 accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta
     5761 cagcttacga accgaacagg cttatgtcca ctgggttcgt gccttcatcc gtttccacgg
     5821 tgtgcgtcac ccggcaacct tgggtagcag cgaagtcgag gcatttctgt cctggctggt
     5881 catgaccaaa atcccttaac gtgagtcagc ctgccgcctt gggccgggtg atgtcgtact
     5941 tgcccgccgc gaactcggtt accgtccagc ccagcgcgac cagctccggc aacgcctcgc
     6001 gcacccgctt gcggcgcttg cgcatggtcg aaccactggc ctctgacggc cagacatagc
     6061 cgcacaaggt atctatggaa gccttgccgg ttttgccggg gtcgatccag ccacacagcc
     6121 gctggtgcag caggcgggcg gtttcgctgt ccagcgcccg cacctcgtcc atgctgatgc
     6181 gcacatgctg gccgccaccc atgacggcct gcgcgatcaa ggggttcagg gccacgtaca
     6241 ggcgcccgtc cgcctcgtcg ctggcgtact ccgacagcag ccgaaacccc tgccgcttgc
     6301 ggccattctg ggcgatgatg gataccttcc aaaggcgctc gatgcagtcc tgtatgtgct
     6361 tgagcgcccc accactatcg acctctgccc cgatttcctt tgccagcgcc cgatagctac
     6421 ctttgaccac atggcattca gcggtgacgg cctcccactt gggttccagg aacagccgga
     6481 gctgccgtcc gccttcggtc ttgggttccg ggccaagcac taggccatta ggcccagcca
     6541 tggccaccag cccttgcagg atgcgcagat catcagcgcc cagcggctcc gggccgctga
     6601 actcgatccg cttgccgtcg ccgtagtcat acgtcacgtc cagcttgctg cgcttgcgct
     6661 cgccccgctt gagggcacgg aacaggccgg gggccagaca gtgcgccggg tcgtgccgga
     6721 cgtggctgag gctgtgcttg ttcttaggct tcaccacggg gcaccccctt gctcttgcgc
     6781 tgcctctcca gcacggcggg cttgagcacc ccgccgtcat gccgcctgaa ccaccgatca
     6841 gcgaacggtg cgccatagtt ggccttgctc acaccgaagc ggacgaagaa ccggcgctgg
     6901 tcgtcgtcca caccccattc ctcggcctcg gcgctggtca tgctcgacag gtaggactgc
     6961 cagcggatgt tatcgaccag taccgagctg ccccggctgg cctgctgctg gtcgcctgcg
     7021 cccatcatgg ccgcgccctt gctggcatgg tgcaggaaca cgatagagca cccggtatcg
     7081 gcggcgatgg cctccatgcg accgatgacc tgggccatgg ggccgctggc gttttcttcc
     7141 tcgatgtgga accggcgcag cgtgtccagc accatcaggc ggcggccctc ggcggcgcgc
     7201 ttgaggccgt cgaaccactc cggggccatg atgttgggca ggctgccgat cagcggctgg
     7261 atcagcaggc cgtcagccac ggcttgccgt tcctcggcgc tgaggtgcgc cccaagggcg
     7321 tgcaggcggt gatgaatggc ggtgggcggg tcttcggcgg gcaggtagat caccgggccg
     7381 gtgggcagtt cgcccacctc cagcagatcc ggcccgcctg caatctgtgc ggccagttgc
     7441 agggccagca tggatttacc ggcaccaccg ggcgacacca gcgccccgac cgtaccggcc
     7501 accatgttgg gcaaaacgta gtccagcggt ggcggcgctg ctgcgaacgc ctccagaata
     7561 ttgataggct tatgggtagc cattgattgc ctcctttgca ggcagttggt ggttaggcgc
     7621 tggcggggtc actacccccg ccctgcgccg ctctgagttc ttccaggcac tcgcgcagcg
     7681 cctcgtattc gtcgtcggtc agccagaact tgcgctgacg catccctttg gccttcatgc
     7741 gctcggcata tcgcgcttgg cgtacagcgt cagggctggc cagcaggtcg ccggtctgct
     7801 tgtccttttg gtctttcata tcagtcaccg agaaacttgc cggggccgaa aggcttgtct
     7861 tcgcggaaca aggacaaggt gcagccgtca aggttaaggc tggccatatc agcgactgaa
     7921 aagcggccag cctcggcctt gtttgacgta taaccaaagc caccgggcaa ccaatagccc
     7981 ttgtcacttt tgatcaggta gaccgaccct gaagcgcttt tttcgtattc cataaaaccc
     8041 ccttctgtgc gtgagtactc atagtataac aggcgtgagt accaacgcaa gcactacatg
//
