LOCUS       Exported                5269 bp ds-DNA     circular SYN 28-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5269)
  AUTHORS   Brian Pfleger Group
  TITLE     Direct Submission
  JOURNAL   Exported Feb 5, 2018 from SnapGene 4.1.5
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5269
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    17..22
                     /label=AvrII
                     /note="Restriction site used by the Bgl-brick team to 
                     define the END of the inducible expression casette, which 
                     spans from the repressor up to the terminator of cds."
     misc_feature    23..121
                     /label=gBlocks part 2
     terminator      23..112
                     /label=ECK120029600
                     /note="Voigt's strongest tested natural promoter"
     primer_bind     23..45
                     /label=3.R
     primer_bind     complement(91..122)
                     /label=WN160
     primer_bind     94..143
                     /label=JMR493
                     /note="araE F :: MS const"
     primer_bind     complement(94..122)
                     /label=3.F
     primer_bind     complement(94..121)
                     /label=JMR491
                     /note="MS const F :: araE"
     misc_feature    113..121
                     /label=Spacer
                     /note="Spacer as is found in the construct used by Voigt 
                     team to test their terminator library."
     gene            complement(122..1540)
                     /gene="araE"
                     /gene_synonym="ECK2839; JW2809"
                     /locus_tag="b2841"
                     /label=araE
                     /db_xref="EcoGene:EG10056"
                     /db_xref="GeneID:947341"
     CDS             complement(122..1540)
                     /codon_start=1
                     /transl_table=11
                     /gene="araE"
                     /gene_synonym="ECK2839; JW2809"
                     /locus_tag="b2841"
                     /product="arabinose transporter"
                     /function="transport; Transport of small molecules: 
                     Carbohydrates, organic acids, alcohols"
                     /label=araE
                     /note="low-affinity L-arabinose transport system proton 
                     symport protein"
                     /db_xref="GI:16130745"
                     /db_xref="ASAP:ABE-0009319"
                     /db_xref="UniProtKB/Swiss-Prot:P0AE24"
                     /db_xref="EcoGene:EG10056"
                     /db_xref="GeneID:947341"
                     /protein_id="NP_417318.1"
                     /translation="MVTINTESALTPRSLRDTRRMNMFVSVAAAVAGLLFGLDIGVIAG
                     ALPFITDHFVLTSRLQEWVVSSMMLGAAIGALFNGWLSFRLGRKYSLMAGAILFVLGSI
                     GSAFATSVEMLIAARVVLGIAVGIASYTAPLYLSEMASENVRGKMISMYQLMVTLGIVL
                     AFLSDTAFSYSGNWRAMLGVLALPAVLLIILVVFLPNSPRWLAEKGRHIEAEEVLRMLR
                     DTSEKAREELNEIRESLKLKQGGWALFKINRNVRRAVFLGMLLQAMQQFTGMNIIMYYA
                     PRIFKMAGFTTTEQQMIATLVVGLTFMFATFIAVFTVDKAGRKPALKIGFSVMALGTLV
                     LGYCLMQFDNGTASSGLSWLSVGMTMMCIAGYAMSAAPVVWILCSEIQPLKCRDFGITC
                     STTTNWVSNMIIGATFLTLLDSIGAAGTFWLYTALNIAFVGITFWLIPETKNVTLEHIE
                     RKLMAGEKLRNIGV"
     primer_bind     complement(1515..1572)
                     /label=JMR492
                     /note="araE :: MS const R"
     primer_bind     1538..1569
                     /label=WN233
     misc_feature    1552..1721
                     /label=gBlocks part 1
     primer_bind     1552..1575
                     /label=JMR490
                     /note="MS const R :: araE"
     misc_feature    1552..1554
                     /label=RBS
     misc_feature    1572..1605
                     /label=HP4
     primer_bind     complement(1578..1605)
                     /label=rTBC0090
     primer_bind     complement(1578..1605)
                     /label=rTBC0088
     primer_bind     complement(1578..1605)
                     /label=rTBC0086
     primer_bind     complement(1578..1605)
                     /label=rTBC0084
     primer_bind     complement(1584..1605)
                     /label=WN167
     primer_bind     1584..1605
                     /label=WN162
     primer_bind     1605..1630
                     /label=WN231
     misc_feature    1605
                     /label=TSS
     promoter        1606..1640
                     /label=BBa_J23101
     -10_signal      1612..1616
                     /label=-10
     primer_bind     1618..1640
                     /label=WN164
     -35_signal      1635..1640
                     /label=-35
     primer_bind     1639..1670
                     /label=rTBC0089
     primer_bind     1639..1670
                     /label=rTBC0087
     primer_bind     1639..1670
                     /label=rTBC0085
     primer_bind     1641..1670
                     /label=rTBC0091
     primer_bind     1641..1670
                     /label=WN168
     primer_bind     complement(1641..1670)
                     /label=WN161
     misc_feature    1641..1660
                     /label=AT-rich random 20-bp
     misc_feature    1661..1721
                     /label=L3S2P21
                     /note="Voigt's strongest tested synthetic promoter"
     primer_bind     1661..1682
                     /label=WN165
     primer_bind     complement(1661..1682)
                     /label=WN163
     primer_bind     1708..1725
                     /label=rTBC0162
     misc_feature    1722..1727
                     /label=AatII
                     /note="Restriction site used by the Bgl-brick team to 
                     define the BEGINNING of the inducible expression casette, 
                     which spans from the repressor up to the terminator of 
                     cds."
     promoter        1728..1768
                     /label=NEOKAN_promoter
     primer_bind     1748..1789
                     /label=JMR496
                     /note="pBb(SEVA224)1g-GFP :: araE R"
     primer_bind     complement(1748..1768)
                     /label=JMR494
                     /note="NCR5/6 :: gent and RK2 F"
     promoter        1893..1921
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     1893..1911
                     /label=rTBC0251
     primer_bind     1893..1911
                     /label=rTBC0211
     primer_bind     1893..1911
                     /label=rTBC0184
     misc_feature    1893..1898
                     /label=-35 box
                     /note="-35 box"
     misc_feature    1913..1921
                     /label=-10 box
                     /note="-10 box"
     primer_bind     complement(1943..1964)
                     /label=JMR345
     misc_feature    complement(2035..2053)
                     /label=CMJ134
                     /note="CMJ134"
     CDS             2110..2643
                     /codon_start=1
                     /label=gentamicin-3-acetyltransferase
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     2644..2665
                     /label=JMR340
     primer_bind     complement(2677..2695)
                     /label=rTBC0257
     primer_bind     complement(2844..2862)
                     /label=rTBC0188
     primer_bind     complement(2845..2864)
                     /label=rTBC0212
     primer_bind     2874..2925
                     /label=WN131
                     /note="Vector.REV"
     primer_bind     complement(2884..2933)
                     /label=WN129
                     /note="Fragment.REV"
     misc_feature    2904..2909
                     /label=SacI
     terminator      2915..3009
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2991..3033)
                     /label=WN72
     primer_bind     complement(2991..3033)
                     /label=Vector.FOR
     primer_bind     3009..3053
                     /label=WN75
     primer_bind     3009..3053
                     /label=Fragment.REV
     misc_feature    3016..3021
                     /label=SpeI (Pseudo-XbaI)
     primer_bind     complement(3091..3108)
                     /label=rTBC0163
     primer_bind     3103..3122
                     /label=rTBC0170
     rep_origin      3121..3652
                     /label=oriV
                     /note="incP origin of replication"
     primer_bind     complement(3354..3373)
                     /label=WN81
     primer_bind     3481..3501
                     /label=Pseq2
     primer_bind     complement(3909..3929)
                     /label=JMR494
                     /note="NCR5/6 :: gent and RK2 F"
     gene            4045..5193
                     /gene="trfA"
                     /label=trfA
     CDS             4045..5193
                     /codon_start=1
                     /transl_table=11
                     /gene="trfA"
                     /product="replication protein"
                     /label=trfA
                     /db_xref="GI:525567860"
                     /protein_id="AGR55406.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     primer_bind     4177..4197
                     /label=Pseq1
     primer_bind     4998..5020
                     /label=WN84
     primer_bind     complement(5194..5216)
                     /label=rTBC0171
     primer_bind     complement(5227..5250)
                     /label=WN74
     primer_bind     complement(5227..5250)
                     /label=Fragment.FOR
     primer_bind     complement(5232..5269)
                     /label=JMR497
                     /note="pBb(SEVA224)1g-GFP :: araE F"
     misc_feature    5244..5249
                     /label=AvrII
     primer_bind     5251..5269
                     /label=JMR495
                     /note="NCR5/6 :: gent RK2 R"
ORIGIN
        1 agactatcgg ctgtagccta ggttgagaag agaaaagaaa accgccgatc ctgtccaccg
       61 cattactgca aggtagtgga caagaccggc ggtcttaagt tttttggctg aagaattctt
      121 atcagacgcc gatatttctc aacttctcgc ctgccatcag tttgcgttcg atatgttcca
      181 gcgtgacatt tttggtttcc ggaatgagcc agaaagtaat gcccacaaac gcaatgttca
      241 gcgcagtgta gagccagaac gtaccggcag cgccaatgct atcaagcagt gtcaggaagg
      301 tcgcgccgat aatcatattc gacacccagt tcgtggtggt cgaacaggta ataccgaaat
      361 cgcggcattt cagcggctga atttcagagc acaggatcca caccactggc gcggcgctca
      421 tcgcataacc ggcaatacac atcatcgtca tgccaacaga gagccaggac aagccactgg
      481 aagccgtacc gttatcaaac tgcatcaggc aatagcccag caccagagtg cctaacgcca
      541 tcacgctgaa accaattttc agagccggtt tacgccctgc tttatctacc gtaaacaccg
      601 caataaaggt ggcgaacata aaggtcagcc ctacgaccag agtcgcaatc atctgttgtt
      661 ctgtggtcgt aaagcccgcc attttgaaga tacgcggcgc gtagtacatg atgatgttca
      721 taccggtaaa ctgctgcatc gcctgcaaca acataccgag aaacacagca cgacggacgt
      781 tacggttgat cttaaacagt gcccaaccgc cctgttttaa cttcaggctt tcacgaattt
      841 cgttgagttc ttctcgcgct ttttccgacg tatcgcgcag catacgcaat acttcttccg
      901 cctcaatatg acgccccttt tccgccagcc agcgcgggct atttggcagg aagactacca
      961 gaataatcag cagaactgct ggtaaagcaa gaacccccaa cattgcgcgc cagttaccgc
     1021 tataactgaa cgctgtatcg gataaaaacg ccagcacgat gccgagtgtg accatcaact
     1081 ggtacatact gatcatctta ccgcgaacgt tttcacttgc catttcagaa agatacagag
     1141 gagcggtgta agacgcgatc ccgacagcaa tgcccagcac cacacgagcg gcgattaaca
     1201 tctctacgct ggtcgcaaaa gcggacccta tagaaccgag tacaaacagg atggcccccg
     1261 ccatcaggct gtatttacgc cccaggcgga acgacagcca accattaaac agcgcaccaa
     1321 ttgctgcacc gagcatcatg ctactaacca cccattcctg caaacgactg gtcagcacaa
     1381 agtgatcggt aatgaacggc aacgctccgg cgattacgcc gatatcaaga ccaaataaca
     1441 atcctgcgac cgcagcagct accgaaacaa acatattcat acgccgcgta tcccgcaaag
     1501 aacgtggcgt taaagcagat tccgtattga tagtaaccat ggaattaacc tcctacttga
     1561 ctttatgagt tcgtcaacaa tatctcactc gagataagtc gacgtgctag cacagtccct
     1621 aggactgagc tagctgtaaa tatgtatgct agttcaagat ggaccaaaac gaaaaaaggc
     1681 ccccctttcg ggaggcctct tttctggaat ttggtaccga ggacgtcgga attgccagct
     1741 ggggcgccct ctggtaaggt tgggaagcca tggaagccat cacaaacggc atgatgaacc
     1801 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggacgca
     1861 caccgtggaa acggatgaag gcacgaaccc agttgacata agcctgttcg gttcgtaaac
     1921 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     1981 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta
     2041 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     2101 gcagcaacga tgttacgcag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     2161 aagttaggtg gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc
     2221 aaatccatgc gggctgctct tgatcttttc ggtcgtgagt tcggagacgt agccacctac
     2281 tcccaacatc agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc
     2341 gcgcttgctg ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc
     2401 aggtttgagc agccgcgtag tgagatctat atctatgatc tcgcagtctc cggcgagcac
     2461 cggaggcagg gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt
     2521 ggtgcttatg tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat
     2581 acaaagttgg gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc
     2641 taacaattcg ttcaagccga gatcggcttc ccggccgcgg agttgttcgg taaattgtca
     2701 caacgccgcc aggtggcact tttcggggaa atgtgcgcgc ccgcgttcct gctggcgctg
     2761 ggcctgtttc tggcgctgga cttcccgctg ttccgtcagc agcttttcgc ccacggcctt
     2821 gatgatcgcg gcggccttgg cctgcatatc ccgattcaac ggccccaggg cgtccagaac
     2881 gggcttcagg cgctcccgaa ggtgagctcg cttggactcc tgttgataga tccagtaatg
     2941 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
     3001 ggtgagaatc caagcactag tgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3061 ctgcggggga tcaggaccgc tgccggagcg caacccactc actacagcag agccatgtag
     3121 ggccgccggc gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg
     3181 gacgttgaca cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct
     3241 cgatttcggc cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt
     3301 tacgcgagtt tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga
     3361 cacttgaggg gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag
     3421 agtgctgaca gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa
     3481 tccagcattt gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct
     3541 gcttttaaac caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga
     3601 ccgcgcacgc cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg
     3661 cctcccatcc ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc
     3721 agccacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
     3781 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
     3841 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
     3901 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     3961 gccaaggatc tgatggcgca ggggatcaag atcgacggat cgatccgggg aattaattcc
     4021 ggggcaatcc cgcaaggagg gtgaatgaat cggacgtttg accggaaggc atacaggcaa
     4081 gaactgatcg acgcggggtt ttccgccgag gatgccgaaa ccatcgcaag ccgcaccgtc
     4141 atgcgtgcgc cccgcgaaac cttccagtcc gtcggctcga tagtccagca agctacggcc
     4201 aagatcgagc gcgacagcgt gcaactggct ccccctgccc tgcccgcgcc atcggccgcc
     4261 gtggagcgtt cgcgtcgtct cgaacaggag gcggcaggtt tggcgaagtc gatgaccatc
     4321 gacacgcgag gaactatgac gaccaagaag cgaaaaaccg ccggcgagga cctggcaaaa
     4381 caggtcagcg aagccaagca ggccgcgttg ctgaaacaca cgaagcagca gatcaaggaa
     4441 atgcagcttt ccttgttcga tattgcgccg tggccggaca cgatgcgagc gatgccaaac
     4501 gacacggccc gctctgccct gttcaccacg cgcaacaaga aaatcccgcg cgaggcgctg
     4561 caaaacaagg tcattttcca cgtcaacaag gacgtgaaga tcacctacac cggcgtcgag
     4621 ctgcgggccg acgatgacga actggtgtgg cagcaggtgt tggagtacgc gaagcgcacc
     4681 cctatcggcg agccgatcac cttcacgttc tacgagcttt gccaggacct gggctggtcg
     4741 atcaatggcc ggtattacac gaaggccgag gaatgcctgt cgcgcctaca ggcgacggcg
     4801 atgggcttca cgtccgaccg cgttgggcac ctggaatcgg tgtcgctgct gcaccgcttc
     4861 cgcgtcctgg accgtggcaa gaaaacgtcc cgttgccagg tcctgatcga cgaggaaatc
     4921 gtcgtgctgt ttgctggcga ccactacacg aaattcatat gggagaagta ccgcaagctg
     4981 tcgccgacgg cccgacggat gttcgactat ttcagctcgc accgggagcc gtacccgctc
     5041 aagctggaaa ccttccgcct catgtgcgga tcggattcca cccgcgtgaa gaagtggcgc
     5101 gagcaggtcg gcgaagcctg cgaagagttg cgaggcagcg gcctggtgga acacgcctgg
     5161 gtcaatgatg acctggtgca ttgcaaacgc tagggccttg tggggtcagt tccggctggg
     5221 ggttcagcag ccacctgcat cgccctaggt cccgtaagtg cgctgttcc
//
