LOCUS       Exported                6093 bp DNA     circular SYN 30-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6093)
  AUTHORS   alenavoit
  TITLE     Direct Submission
  JOURNAL   Exported Nov 17, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6093
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     RBS             38..60
                     /label=strong RBS
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             68..3058
                     /codon_start=1
                     /product="Cellodextrin Phosphorylase from Clostridium 
                     cellulosi"
                     /label=cellodextrin phosphorylase
                     /translation="MRGSHHHHHHGSITKVTPRSCKLTPFEILKQKFGSIIAMSGCADA
                     IYSDSSCKNVIGITGLPIKAPVIQVLLENSKLAAFLNEYVQNKSVCLALEGMRYCLVFE
                     DGQKVKLPQELDDIIQSTVSDVKNWAGTLTENGEHVIDLKKPSPGPHFYVNLLIGNRLG
                     FNRTLQTTPKSVVDRFGRGSFRSHAATQVLATRYDMRQEENGFPANRQFYLYENGKQIF
                     YSGSVDDDNILEAKCIHSCNRTVIKYKTACKLEITRTIFLCPQKKGLPLATEIQIINIK
                     NNSERKRNLSITYTGMFGTAATHAIFEDVTYTNVIMQSAALYNDNGEFIGITPDYYRKK
                     YNEDTRFVTMFVRKDGKRIFPKSFCTDYNDFVGNGTLEHPAGGEHLNNKLNRKGPGFFA
                     LGTPFTIMPGQDIEIDTFTGMSSSKDNPNYDDTTMPKELEALLRYFEASDSVQSALREV
                     IDFQEKYCKYIEFKTEDKMFNAGFNKNLAFQILYQTFMSRSFCQTQKGYREIGFREIQD
                     LYASMYYFINLGYQDFVKNLLFEWTSNVYKMGYANHNFYWVGKQPGLYSDDSLWLLQAY
                     YRYIIYTKDTSILNEEVPVADGNNEKRAVRDTLKAIIQYSAFISVGNHGLPLLDLADWN
                     DCLKIDNDYVDGAQKEKLYYKQLKETGGKYGDRFMSDYSESVMNAFLLKVAVDDLAEIA
                     ELDKDAELEQSMRKISSALSEKIQKYAWKGNFFARVLLNRYKDGSFTYVGAKGDKLSAD
                     PNIDGTYFLNSFSWAILSGVATDEQIGIMVDTLKKYMLTPYGIRLTTPADLNKLANDTA
                     TDHYFYGDRENGAVFKHASMMATAALAKAAKSVKDKALAKEMARLAYYMIDLVLPYKNL
                     EDPFKLAGNPRICTQYINADSGENIGPLLSGTATWLNLTITSLAGIEYVKDGIMFSPVL
                     REDETRLEYSLKAPGCSYNITITKPRGFARLIDSAYDLRIDGKKTDNCIIPMFTDGKEH
                     SVTLEFK"
     CDS             68..70
                     /codon_start=1
                     /product="start codon"
                     /label=ATG
                     /translation="M"
     CDS             80..97
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             80..97
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(108..130)
                     /label=pBP2600 CcCDP rev
     primer_bind     complement(934..955)
                     /label=pBP2616
     primer_bind     2195..2210
                     /label=pBP2617
     unsure          2294..2296
                     /label=Val (Ile in protein sequence)
     primer_bind     2639..2668
                     /label=pBP2601 CcCDP fw
     primer_bind     complement(3023..3112)
                     /label=pBP2595 T7TT rev
     terminator      3065..3112
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3083..3133)
                     /label= pBP2623 T7TT rev
     primer_bind     complement(3100..3112)
                     /label=pBP2311-Gibson27-Rev
     primer_bind     3122..3164
                     /label=pBP2618 rrnC-KanR rev
     primer_bind     3125..3170
                     /label=BglII-XhoI-KanR rev
     terminator      3131..3160
                     /label=rrnC
     CDS             complement(3167..3961)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase from Tn5confers
                     resistance to neomycin, kanamycin, and G418 (Geneticin)
                     "
                     /label=neokan
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     3341..3367
                     /label=pBP2314
     primer_bind     complement(3374..3427)
                     /label=pBP2302-Gibson23-For
     primer_bind     3374..3427
                     /label=pBP2301-Gibson22-Rev
     primer_bind     3390..3409
                     /label=pBP2284-PCR9
     primer_bind     complement(3394..3410)
                     /label=pBP2273
                     /note="Begin  gBlock pBP2248 For"
     primer_bind     3838..3854
                     /label=pBP2270
                     /note="Neokan-Rev  +100  xSeq"
     primer_bind     complement(3890..3913)
                     /label=pBP2269
                     /note="Neokan-For  +50  xSeq"
     primer_bind     complement(3930..4007)
                     /label=pBP2619 RBS-KanR fw
     misc_feature    3968..3988
                     /label=PART 2
     RBS             3968..3979
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     3973..4007
                     /label=pBP2262 LacI fw
     misc_feature    3983..3988
                     /label=PART 3
     misc_feature    3989..5149
                     /label=PART 3
     CDS             complement(3989..5071)
                     /codon_start=1
                     /label=LacI
                     /label=LacI (Ecogene EG10525)
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3989..4009
                     /label=pBP469
     primer_bind     complement(4065..4080)
                     /label=pBP560
     primer_bind     4765..4782
                     /label=BPp162
                     /note="forward sequencing primer for lacI 
                     		60.4	PCR	Natascha Hotz"
     primer_bind     4811..4827
                     /label=pBP562
     promoter        complement(5072..5149)
                     /label=PlacIq
                     /note="mutant lacIQ promoter; contains an ""up"" mutation 
                     in the -35 box at position -35 of the lacI promoter (Calos 
                     (1978) Nature 274(5673): 762-5)"
     primer_bind     5126..5183
                     /label=pBP2621 p15A fw
     primer_bind     complement(5126..5155)
                     /label=pBP2620 PlacIQ fw
     primer_bind     complement(5126..5155)
                     /label=lacI rev
     primer_bind     5140..5155
                     /label=PBICI fw
     misc_feature    complement(5156..6065)
                     /label=p15a ori
                     /note="Chang and Cohen 1978
                     pACYC
                     "
     primer_bind     5156..5187
                     /label=pBP2298-Gibson21-For
     primer_bind     complement(5156..5187)
                     /label=pBP2311-Gibson27-Rev
     primer_bind     5156..5173
                     /label=pBP2281-PCR8
     primer_bind     complement(5225..5245)
                     /label=pBP2312
     rep_origin      complement(5420..5965)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(join(6032..6093,1..12))
                     /label=pBP2622 p15A rev
     primer_bind     complement(6035..6065)
                     /label=pBP2299-Gibson21-Rev
     primer_bind     complement(6038..6065)
                     /label=pBP2282-PCR8
     primer_bind     join(6069..6093,1..5)
                     /label=PBICI fw
     primer_bind     join(6073..6093,1..12)
                     /label=pBP2594 PT7 fw
     promoter        6073..6091
                     /label=T7_promoter
     protein_bind    join(6092..6093,1..23)
                     /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    join(6092..6093,1..23)
                     /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)."
ORIGIN
        1 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
       61 tatacatatg agaggatcgc atcaccatca ccatcacgga tccattacca aagtgacccc
      121 gcgtagctgc aaactgaccc cgttcgagat tctgaaacaa aaattcggta gcattattgc
      181 gatgagcggc tgcgcggatg cgatctacag cgacagcagc tgcaagaacg tgatcggtat
      241 taccggcctg ccgatcaaag cgccggtgat tcaggttctg ctggagaaca gcaagctggc
      301 ggcgttcctg aacgaatacg tgcaaaacaa aagcgtttgc ctggcgctgg agggtatgcg
      361 ttattgcctg gtgtttgaag acggccagaa ggttaaactg ccgcaagagc tggacgatat
      421 cattcagagc accgtgagcg atgttaaaaa ctgggcgggt accctgaccg agaacggcga
      481 acacgtgatc gacctgaaga aaccgagccc gggtccgcac ttctacgtta acctgctgat
      541 tggtaaccgt ctgggcttta accgtaccct gcaaaccacc ccgaagagcg tggttgatcg
      601 tttcggtcgt ggcagctttc gtagccatgc ggcgacccag gttctggcga cccgttacga
      661 catgcgtcaa gaggaaaacg gcttcccggc gaaccgtcag ttttacctgt atgagaacgg
      721 caagcaaatc ttctacagcg gcagcgtgga cgatgacaac atcctggaag cgaaatgcat
      781 tcacagctgc aaccgtaccg ttattaagta taaaaccgcg tgcaagctgg agatcacccg
      841 taccattttt ctgtgcccgc agaagaaagg tctgccgctg gcgaccgaga tccaaatcat
      901 caacatcaag aacaacagcg aacgtaaacg taacctgagc atcacctaca ccggcatgtt
      961 tggtaccgcg gcgacccacg cgatctttga agatgtgacc tacaccaacg ttattatgca
     1021 gagcgcggcg ctgtataacg acaacggcga gttcatcggc attaccccgg attactatcg
     1081 taagaaatac aacgaagaca cccgtttcgt gaccatgttt gttcgtaagg acggtaaacg
     1141 tatcttcccg aagagcttct gcaccgatta taacgacttt gtgggtaacg gtaccctgga
     1201 gcacccggcg ggtggcgaac acctgaacaa caagctgaac cgtaaaggtc cgggcttctt
     1261 tgcgctgggt accccgttca ccattatgcc gggccaagac atcgagattg atacctttac
     1321 cggtatgagc agcagcaagg ataacccgaa ctacgatgac accaccatgc cgaaagagct
     1381 ggaagcgctg ctgcgttatt tcgaagcgag cgatagcgtg cagagcgcgc tgcgtgaggt
     1441 tatcgacttc caagaaaagt actgcaagta catcgagttc aagaccgaag acaagatgtt
     1501 caacgcgggt tttaacaaaa acctggcgtt tcagatcctg taccaaacct tcatgagccg
     1561 tagcttttgc cagacccaaa agggttatcg tgagatcggc ttccgtgaaa ttcaggatct
     1621 gtacgcgagc atgtactact tcatcaacct gggttaccaa gacttcgtga agaacctgct
     1681 gtttgagtgg accagcaacg tttacaaaat gggctatgcg aaccacaact tctactgggt
     1741 tggtaaacag ccgggcctgt atagcgatga cagcctgtgg ctgctgcaag cgtactatcg
     1801 ttacatcatt tataccaagg ataccagcat cctgaacgag gaagtgccgg ttgcggatgg
     1861 taacaacgaa aagcgtgcgg tgcgtgacac cctgaaagcg atcattcagt acagcgcgtt
     1921 catcagcgtt ggtaaccatg gtctgccgct gctggatctg gcggactgga acgattgcct
     1981 gaagattgat aacgactatg tggacggtgc gcagaaggag aaactgtact ataagcaact
     2041 gaaagaaacc ggtggcaaat acggcgatcg tttcatgagc gactatagcg agagcgttat
     2101 gaacgcgttt ctgctgaagg tggcggttga tgacctggcg gagatcgcgg aactggataa
     2161 ggacgcggag ctggaacaga gcatgcgtaa aatcagcagc gcgctgagcg aaaagattca
     2221 aaaatacgcg tggaagggta acttctttgc gcgtgtgctg ctgaaccgtt acaaagatgg
     2281 cagcttcacc tatgttggtg cgaagggcga caaactgagc gcggacccga acatcgatgg
     2341 tacctatttc ctgaacagct ttagctgggc gattctgagc ggtgtggcga ccgatgagca
     2401 gatcggcatt atggttgaca ccctgaagaa atacatgctg accccgtatg gcatccgtct
     2461 gaccaccccg gcggatctga acaaactggc gaacgacacc gcgaccgatc actacttcta
     2521 tggtgaccgt gaaaacggcg cggtgtttaa gcatgcgagc atgatggcga ccgcggcgct
     2581 ggcgaaggcg gcgaaaagcg ttaaggataa agcgctggcg aaagagatgg cgcgtctggc
     2641 gtactatatg atcgatctgg tgctgccgta caagaacctg gaagacccgt tcaaactggc
     2701 gggtaacccg cgtatctgca cccagtatat taacgcggac agcggcgaga acattggtcc
     2761 gctgctgagc ggtaccgcga cctggctgaa cctgaccatc accagcctgg cgggtattga
     2821 gtacgtgaag gatggcatta tgttcagccc ggttctgcgt gaggacgaaa cccgtctgga
     2881 atacagcctg aaggcgccgg gttgcagcta taacatcacc attaccaaac cgcgtggctt
     2941 tgcgcgtctg atcgacagcg cgtacgatct gcgtattgac ggtaagaaaa ccgacaactg
     3001 cattatcccg atgttcaccg atggtaaaga gcatagcgtg accctggagt tcaagtaaga
     3061 cgtcctagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgaagcttat
     3121 aatactcgag taaaaaaaat ccttagcttt cgctaaggat gtgcactcag aagaactcgt
     3181 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     3241 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     3301 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     3361 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc
     3421 cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     3481 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
     3541 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     3601 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     3661 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     3721 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct
     3781 ggagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     3841 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     3901 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
     3961 tgtcgacttt ctcctcttta atagatcttc actgcccgct ttccagtcgg gaaacctgtc
     4021 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4081 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     4141 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     4201 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     4261 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     4321 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     4381 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     4441 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     4501 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     4561 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     4621 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     4681 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     4741 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     4801 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     4861 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     4921 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     4981 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     5041 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     5101 ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcg aattcctagc
     5161 ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt
     5221 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc
     5281 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct
     5341 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
     5401 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
     5461 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5521 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
     5581 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
     5641 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
     5701 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
     5761 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
     5821 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
     5881 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat
     5941 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta
     6001 gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa
     6061 gttgtctgca gttaatacga ctcactatag ggg
//
