LOCUS       pAC11        8497 bp DNA     circular SYN 21-NOV-2022
DEFINITION  synthetic circular DNA
ACCESSION   urn.local...a6-82xqi3h
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8497)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 24, 2023 from SnapGene 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..98
                     /label=ConLS
     primer_bind     1..20
                     /label=AC_09
     primer_bind     complement(33..52)
                     /label=AC_019
     misc_feature    106..130
                     /label=ConLS scar
     misc_feature    135..204
                     /label=PA3
     primer_bind     205..234
                     /label=AC_042
     RBS             215..222
     misc_feature    231..947
                     /label=E2-crimson
     terminator      952..1034
                     /label=rpoC
     primer_bind     complement(1044..1104)
                     /label=AC_041
     CDS             complement(1105..2115)
                     /label=SpecR
     primer_bind     complement(1116..1135)
                     /label=AC_07
     promoter        complement(2116..2249)
                     /label=SpecR Promoter
     primer_bind     complement(2251..2273)
                     /label=AC_10
     primer_bind     2251..2273
                     /label=AC_018
     primer_bind     2254..2293
                     /label=AC_051
     misc_feature    2274..8497
                     /label=pTF-FC2 replicon
     misc_feature    2408..2737
                     /label=oriV
     repeat_region   2538..2552
     repeat_region   2557..2573
     repeat_region   2585..2591
     repeat_region   2597..2603
     repeat_region   2664..2685
     repeat_region   2686..2707
     CDS             complement(2694..3593)
                     /codon_start=1
                     /transl_table=11
                     /gene="repC"
                     /product="replication protein C"
                     /label=repC
                     /protein_id="AAA27382.1"
                     /translation="MEQEKKGESAMSYDLTHARHDPAHCLTPGLFRSLKRGERKRLKLD
                     VTYNYGDDSIRFWGPEPLGGDDLRVLQGLVAMAAISGDNGRGIVLRHETESEAGQQLRL
                     WLDMRWDAIEKDTMVAKGSFRQLARELGYAEDGGSQFKTIRESIERLWAVSVIVERGGK
                     RQGFRILSEYASDEQEGKLFVALNPRLADAVMGERPHTRINMAEVRKLETDPARLLHQR
                     LCGWIDPGKSGKAEIDTLCGYVWPDAANDEAMKKRRQTARKALVELAAVGWTVNEYAKG
                     KWEISRPNPRRNVPQPPA"
     gene            complement(2694..3593)
                     /gene="repC"
                     /label=repC
     repeat_region   complement(2708..2729)
                     /gene="repC"
     primer_bind     complement(3541..3560)
                     /label=AC_056
     CDS             complement(3553..4425)
                     /codon_start=1
                     /transl_table=11
                     /gene="repA"
                     /product="replication protein A"
                     /label=repA
                     /protein_id="AAA27381.1"
                     /translation="MALDIMAAFTNEPPELDFIWPGFLAGTVGALVAPGATGKSFFALE
                     AAMSIACSVAGGDLVGLTPAHTGRVVYLAGEDPQPALVRRVHAIGQHLNQSAREAIAEN
                     LMLEPIMGKRLNVMDEAHLRRVIDYSAGARLIVLDTLSRIHSLDENSNGDMAHLVSVLE
                     HIAATTGAAVLYLHHVNKGSARDGQTDQQQAARGASALIDNARWCGYVAKMTEQEAERM
                     SDRGFDRSPIGNERRGLFVRFGVSKQNYDATPLDRWYQRHSGGVLLPVELQEAISNGAG
                     KKGGKRNEL"
     gene            complement(3553..4425)
                     /gene="repA"
                     /label=repA
     primer_bind     complement(4276..4295)
                     /label=AC_055
     CDS             complement(4491..4706)
                     /codon_start=1
                     /label=pasC / ORF5
                     /translation="MIEYSYQIDPRPSDLGGGWRLRLLESGEEVGGGVFPLSEYATAEN
                     AEEAASYAYEDALAEASAWLASRGEN"
     CDS             complement(4498..4509)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(4706..4975)
                     /codon_start=1
                     /transl_table=11
                     /label=pasB / ORF4
                     /translation="MAWKVELDPAAERELGKIDQQTARRILAFLHGRVAQLDDPRSIGE
                     ALKGSKLGAFWKYRVGDWRIIASIEDGALRILVMRIGNRKEVYRQ"
     RBS             4716..4724
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(4959..5183)
                     /codon_start=1
                     /transl_table=11
                     /label=pasA / ORF3
                     /translation="MLAIRLPAEVETRLEALAQATGRTKTFYAREAILEHLDDLEDLYL
                     AEQRLIDIRAGKTQTVPLEEVMKRYGMEG"
     primer_bind     complement(5172..5191)
                     /label=AC_054
     CDS             complement(5240..6301)
                     /codon_start=1
                     /transl_table=11
                     /gene="repB"
                     /product="replication primase"
                     /label=repB
                     /protein_id="AAA27386.1"
                     /translation="MKSEQLKQFERYAEAVGAERYRVTSIKMQADGRKQTFILDKKDGI
                     TRGFTPQEIEQRTPEMQRLQRRGENLYYTPLSDKKHHILIDDMNREKLERLIKDGYQPA
                     AVLESSPGNYQAIITVSKLGTAHDKDVGNRLSDALNREYGDPKLSGAIHPHRAPGYENR
                     KPKHQREDGSYPEVRLLKAERRECVKALALSSQIDAEYQRQAALKAQQPERSKAKPALE
                     LAAASGSAIDAYQRHYRDVLKRQRGGEVDLSRVDSMIAVRMRVTGHDQAAIEGAIRQCA
                     PATRQKDEGRDWNDYAQRTARYAYSAAGDRQAADLGKYRQQWEKLEGREPVRQQEQAKA
                     QKIERDNSPGMSL"
     primer_bind     6004..6023
                     /label=AC_049
     primer_bind     complement(6004..6023)
                     /label=AC_050
     regulatory      6313..6318
                     /gene="repB"
                     /regulatory_class="ribosome_binding_site"
     CDS             complement(6512..7738)
                     /codon_start=1
                     /transl_table=11
                     /gene="mobA"
                     /label=mobA
                     /note="ORF6"
                     /protein_id="AAA27389.1"
                     /translation="MIALSQEAVRSKDTINHYVLSWREGEQPSPEQVEEAVSIFMDELG
                     VKDHQAIYGLHADTDNLHLHLAINRVHPETLKVVKINNGFDIEAAHKAIARIENAQGWQ
                     REQNGRYQVLENGELGREHIDKDKPRQPAQPKRDMENRTGEKSAERIAIEDGAPIIKKA
                     QTWEQLHRELAAKGMRYEKTGSGATLFVGDVGVKASSADRDASLSKLQKRLGAYQPPQR
                     QQVAQREPEPIKPDVPGWKDYITGRKAHYSEKNADKLVQDKRQEQERKQLAEQQKARRD
                     ELMRGNWKGKGEVLNAMRSVIAAEQAAEKAALKEKHQKQREQHRQQFRPYPDLEQWQRM
                     QKSPELAEQWRHRASEPQRIEGDRSEPPTPRDIRAYQPEIVGQQVHYSRKEEGGRGGGV
                     SFVDKGKSID"
     gene            complement(6512..7738)
                     /gene="mobA"
                     /label=mobA
     primer_bind     complement(6781..6800)
                     /label=AC_053
     primer_bind     complement(7666..7685)
                     /label=AC_052
     oriT            8351..8405
     primer_bind     complement(join(8478..8497,1..20))
                     /label=AC_048
ORIGIN
        1 atctgaatca tgcgcggatg acacgaactc acgacggcga tcacagacat taacccacag
       61 tacagacact gcgacaacgt ggcaattcgt cgcaataccg tctcactgaa ctggccgata
      121 attgcagacg aacgggtgaa acaaaacggt tgacaacatg aagtaaacac ggtacgatgt
      181 accacatgaa acgacagtga gtcaatacag aaacagagga gatattacat atggatagca
      241 ctgagaacgt catcaagccc ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
      301 gccacgagtt cgagatcgag ggcgtgggcg agggcaagcc ctacgagggc acccagaccg
      361 ccaagctgca agtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
      421 agttcttcta cggctccaag gcgtacatca agcaccccgc cgacatcccc gactacctca
      481 agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
      541 tggtgaccgt gacccaggac tcctccctgc aggacggcac cctcatctac cacgtgaagt
      601 tcatcggcgt gaacttcccc tccgacggcc ccgtaatgca gaagaagact ctgggctggg
      661 agccctccac tgagcgcaac tacccccgcg acggcgtgct gaagggcgag aaccacatgg
      721 cgctgaagct gaagggcggc ggccactacc tgtgtgagtt caagtccatc tacatggcca
      781 agaagcccgt gaagctgccc ggctaccact acgtggacta caagctcgac atcacctccc
      841 acaacgagga ctacaccgtg gtggagcagt acgagcgcgc cgaggcccgc caccacctgt
      901 tccagactca cggtatggac gaattgtaca agcacgacga attgtaaatc cgtaatcgtt
      961 aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga
     1021 aagagcattt gtcagctgga tcagcagatg ggaaccggat actcctcagc ctccagagta
     1081 gccaatgaga cgccgagcgg ccgcttattt gccgactacc ttggtgatct cgcctttcac
     1141 gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc
     1201 aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat
     1261 tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat
     1321 gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca
     1381 tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag
     1441 ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat
     1501 agccagatca atgtcgatcg tggctggctc gaagataccc gcaagaatgt cattgcgctg
     1561 ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg
     1621 cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt
     1681 ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt
     1741 aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa
     1801 atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag
     1861 ttgagtcgat acttcggcga tcaccgcttc cctcataatg tttaactttg ttttagggcg
     1921 actgccctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     1981 acgcgcttgc tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca
     2041 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     2101 gttgcgtgag cgcatacgct acttgcatta cagcttacga accgaacagg cttatgtcca
     2161 ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggtagcag
     2221 cgaagtcgag gcatttctgt cctggctggt catgaccaaa atcccttaac gtgctcaagg
     2281 cagccagaac atccaaaaat ccgctgtagg cgattctgat ggcttctgtt cgggtatgag
     2341 gcgatgtgct ccacaagcac gaccgacaag ccagccgaaa cgctcgccaa gctggccgct
     2401 gcgctacctg ccggagaggc caaacgctac cgggtgatgg tgtccattgt cccggtggaa
     2461 gaatgaaggc gagcggggcg gggcggggtg tgtctggccg ggcttgtccc ggccttgtct
     2521 tttggggttt ttggtaagga cttttgacgc cctcaagggc ggcgaaaagt accgcgccta
     2581 gtggatgatc ttggatagat catggaggga ttttgcgaat cgcccgcaaa cccgcgccgt
     2641 tgctaggttt ttcagatttc ggcggggaac gttacgccgg gggttgggga acgttacgcc
     2701 gggggttggg gaacgttacg ccgggggttg ggcctgctga tttcccactt gcccttggcg
     2761 tactcgttca ccgtccaacc aacggcggca agctcgacaa gcgccttgcg ggcggtctgg
     2821 cggcgctttt tcattgcttc atcgttggct gcgtctggcc atacataacc gcacagcgtg
     2881 tcgatttcag ctttgccaga ctttccgggg tcaatccagc cacatagccg ctggtgtagc
     2941 agccgtgccg ggtctgtttc cagcttgcga acttctgcca tgttgatgcg ggtgtgcggg
     3001 cgctctccca taaccgcatc cgccagccgg ggattaagcg caacaaataa cttgccttct
     3061 tgctcgtcgc tcgcgtactc ggacagaatg cggaaccctt gccgcttacc acctctttcg
     3121 acaatcaccg ataccgccca aagccgttcg atgctttccc ggatggtttt aaactgactt
     3181 cctccatctt cggcgtagcc aagttctcgg gccaactggc ggaagctgcc cttggctacc
     3241 atcgtatctt tctctatggc gtcccaccgc atatcaagcc ataggcggag ttgctggcct
     3301 gcttctgatt ccgtttcgtg ccgtagcacg atgccgcgcc cgttatctcc ggaaattgca
     3361 gccattgcca ccagcccttg caatacgcgc aagtcatcgc cgccaagtgg ttcaggcccc
     3421 caaaaacgga ttgagtcatc tccgtagttg taggtcacat cgagcttgag cctctttcgt
     3481 tctccgcgct tgagactgcg gaaaagcccc ggcgtgaggc aatgcgcggg gtcgtgccgc
     3541 gcatgggtga ggtcatagct cattgcgctt tccccctttt tttcctgctc cattgctgat
     3601 tgcctcctgt agttcaacgg gcaacaacac cccgccgctg tgccgctgat accagcggtc
     3661 tagcggggtc gcgtcgtagt tctgcttgct cacgccaaag cggacaaaaa ggccgcgccg
     3721 ctcgttgccg atgggcgaac gatcaaagcc ccggtcactc atgcgctcgg cttcctgctc
     3781 cgtcattttg gcgacatagc cgcaccatct ggcgttgtcg atcagggcag acgcgccccg
     3841 cgctgcctgc tgctggtcgg tctggccgtc gcgggcgctg cccttgttga cgtggtgcag
     3901 gtacaggaca gccgccccgg tggtcgccgc gatgtgttcc aacacggaaa caaggtgggc
     3961 catgtcgcca ttgctgttct cgtcgaggct gtgaatccgg ctcagggtgt ccagcacaat
     4021 cagccgggcc cctgcgctgt agtcgatgac gcggcgcaag tgcgcctcgt ccatcacgtt
     4081 tagccgcttg cccatgatcg gctcaagcat caggttctca gcgatggctt cgcgggccga
     4141 ctggttgagg tgctggccga tggcgtggac acgtcgcaca agggcgggct gtggatcttc
     4201 gccagcgaga taaaccacgc gcccggtgtg cgccggggtt agtcccacaa ggtcgccgcc
     4261 tgccacactg caagcgattg acatggccgc ttcaagagca aaaaagctct tgccagttgc
     4321 gccaggtgcg acaagtgcgc ccacggttcc ggccaagaat ccgggccaga tgaaatcaag
     4381 ttctggcggc tcattggtga aggccgccat aatgtctaaa gccatgcttg tctccttttc
     4441 agacagttgt ggttaggcgc tgccggggcc gcaatcccct gccgcgccgc tcaattttcg
     4501 cccctcgatg ccagccacgc cgaagcctcg gccaaggcgt cctcataggc gtagctggcc
     4561 gcttcttctg cgttctctgc tgtggcgtac tcggacaacg ggaacactcc gccgccgact
     4621 tcctcgccgc tttccaacag gcgcaaccgc cagccgccgc caaggtcgga ggggcgcggg
     4681 tcgatctggt agctgtattc gatcattggc ggtaaacctc cttacgattg ccgatgcgca
     4741 taacgaggat gcgcaaagca ccgtcctcga tgctggcgat gattcgccaa tccccaacgc
     4801 ggtatttcca gaaggctccc agtttggagc ctttgagggc ttcgccaatg ctgcgcgggt
     4861 cgtcgagctg ggcgacacgg ccatgcaaaa aagcgaggat gcggcgggcg gtctgctggt
     4921 cgatcttgcc tagctcgcgc tcggcggctg ggtcgagttc aaccttccat gccatagcgt
     4981 ttcatcactt cttcgagtgg cacggtttgg gttttgcctg cgcgaatgtc gatcaggcgt
     5041 tgctctgcaa ggtacaaatc ttcgaggtca tccaagtgct caaggatggc ttcgcgggca
     5101 tagaaagtct tggtccgccc tgtggcctgc gccagtgctt caaggcgggt ttccacttcg
     5161 gcgggcagtc ggattgcaag catgtttcgg ctcctgttct gaaaacgcta tacaagtata
     5221 tcacaaccac cacgcaacgc tagagactca ttcccggcga gttgtcgcgc tcgatcttct
     5281 gcgcctttgc ctgctcctgc tgtcgtacag gctcgcgccc ttccagcttc tcccactgct
     5341 gccggtactt gccaagatcg gcggcttgcc ggtcgcctgc ggcgctgtag gcatagcggg
     5401 cggtgcgctg cgcgtagtcg ttccaatcgc ggccctcgtc tttctgccgg gtggccggtg
     5461 cgcactggcg gatagcgccc tcgatggccg cttgatcgtg gccggtgacg cgcatacgca
     5521 cggcaatcat ggaatccacg cgggacaagt ccacctcgcc gccacgctgc cgcttgagca
     5581 cgtcgcggta atgccgctgg taggcgtcga tcgcgctgcc gctggccgct gcaagctcca
     5641 aggcgggctt ggctttgctg cgctcgggct gctgcgcctt caaggccgct tgccgctgat
     5701 actcggcgtc gatctggctg gacaaggcca gcgccttgac gcactcccgg cgctcggcct
     5761 tgagcaagcg cacctctgga tagctgccgt cctcccgctg gtgcttgggc ttgcggttct
     5821 cgtagccggg tgcgcggtgc gggtggatgg ctcccgatag cttcgggtcg ccgtattcac
     5881 ggttcagggc atcgctcagg cggttgccca cgtccttatc gtgggccgtc cccagcttcg
     5941 acacggtgat gatggcctga tagttgccgg ggctggattc cagcacggcg gcgggctggt
     6001 agccgtcttt gataagccgc tccagcttct cccggttcat gtcgtcgatg aggatgtgat
     6061 gcttcttgtc cgatagcggc gtgtagtaga ggttttcgcc ccggcgctgt aggcgctgca
     6121 tctccggcgt gcgctgctcg atctcctgcg gtgtgaaacc ccgcgtgatg ccgtccttct
     6181 tgtcgaggat gaaggtttgc ttccttccgt ctgcctgcat cttgatggac gttacccggt
     6241 agcgctccgc gcctaccgct tccgcgtacc gctcgaactg ctttagctgc tccgatttca
     6301 tcgcctgcgc cctctcctgc tgtatccgct gccgctcctg ctggatgctc tcctgaagct
     6361 ctgcatttgt gatcttgaag ccgtgctctg ctgctaactt cccgcacatg gccttgtatt
     6421 cgtcgttccc ggttacggtg aagctgcccc acttctgcgc cgatagctgc aaggcggcaa
     6481 gcgtgctgtc ccggttgcgc cagtcgtgaa tatcgatgct tttgcccttg tccacgaaag
     6541 acacaccccc gccccgcccc ccttcttcct tccggctgta atgcacctgc tggcccacga
     6601 tttcaggctg ataggcgcga atgtcgcgcg gcgtcggcgg ctcgctgcgg tcgccctcga
     6661 tgcgctgcgg ctcgctggcc cggtgtcgcc actgctcggc aagctccggg cttttctgca
     6721 tccgctgcca ctgctccagg tcgggatagg gtctgaactg ctgccggtgc tgctcgcgct
     6781 gcttctggtg cttctccttc aaggcggctt tctctgcggc ctgctcggcg gcaatcacgc
     6841 tgcgcatggc gttgagcact tcgcccttgc ctttccagtt gccgcgcatg agttcatcgc
     6901 ggcgggcctt ctgctgctcg gcaagttgct tgcgctcctg ctcctgccgc ttgtcctgta
     6961 ccagcttgtc cgcgttcttc tccgagtagt gcgccttgcg cccggtgatg tagtccttcc
     7021 atcccggcac gtctggcttg atcggctccg gctcgcgctg tgctacctgc tgacgctgcg
     7081 gcggctggta agctccgagt cgcttttgca acttggatag gctcgcgtcc cggtctgcgc
     7141 tgctggcctt cacgccaaca tcaccgacaa agagcgtcgc cccgctgcct gttttctcgt
     7201 agcgcatccc cttggctgct aactcgcggt gcaactgctc ccacgtctgc gcctttttga
     7261 tgatcggtgc gccgtcctca atggcgatgc gttcggcgga tttctccccg gtgcggtttt
     7321 ccatgtcgcg cttcggctgg gctggctggc gcggcttgtc cttgtcgatg tgctcccggc
     7381 ctaactcacc attctccaac acctgataac gcccgttctg ctcacgctgc cagccttgtg
     7441 cgttctcgat tcgtgctatc gccttgtgcg ctgcctcaat gtcgaagccg ttattgatct
     7501 tcaccacctt gagcgtttca ggatgcacgc ggttgatggc aaggtgcagg tgcaagttgt
     7561 ccgtgtcggc gtgcaaacca tagatggctt gatggtcttt aacacctagc tcatccatga
     7621 aaatgcttac cgcttcttcg acctgctccg ggctgggctg ctcgccttct cgccaactca
     7681 ggacatagtg attgatcgta tccttgcttc gcactgcctc ttgtgacaag gcaatcattt
     7741 ctgccgtctg gctctgtggt tcatcggtga tgaatccacg cgccccggca tggatgcact
     7801 tctcctgcga gttctcacgc tcaggctcac ggatgtagcc agttaggcgg gaaacgcgaa
     7861 ccgccttgct tgctgccttc tgcgggttct taaccttctt gacgatcacg gctcagtttc
     7921 ccgatatagg ctttcaagac cacaagcgcc ccggctgtgt ccttgctgta cacgcctccg
     7981 ctctcgttgt ggatgtgctt aagcaatcca ccgatgcggc gcaactcttt gagcattacc
     8041 gcgtcggcgc tggcaatgat cggcctgcca aagtagcggc ggcggacaag ctccgacatg
     8101 ctcaacccgg caaggtcggc atcctctttc aagcgggcct tctccgctgc ggtcagccgt
     8161 acattgataa cggcgtccag cggttccggc ccctgcgttt caaatggcat cgtgtctcct
     8221 tctcgcgggg ggcgggcggg gtgtgtcttg gacagtaaga agcgagagcg ccgcactgtc
     8281 cggggtcaag gggatacccc ttgcaggata agccagaccg tgagaagcga agcgccgaat
     8341 gggcaggcgt ccgttggaac cacgtggcga agccatgtgt gttacaacgg tcatcctgta
     8401 ttgctcaacc gctctactat catatcagca gtttaggcac actagatgca ctttgcaaca
     8461 agcataaaag gcgctatgct gtcaactccg aaaccag
//
