LOCUS       Exported                6332 bp ds-DNA     circular SYN 10-SEP-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6332)
  AUTHORS   Ethan
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2018 from SnapGene Viewer 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6332
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..431
                     /label=PMD19-1
     misc_feature    146..431
                     /label=LacZa-1
     misc_feature    515..560
                     /label=CaMV_minimal_promoter
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=#ffff66
                     ApEinfo_fwdcolor=#ffff66"
     misc_feature    576..1310
                     /label=PMDC107-GFP
     CDS             1290..1307
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1351..1364
                     /label=nexteraA
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=#ff8000
                     ApEinfo_fwdcolor=#ff8000"
     misc_feature    1365..1382
                     /label=transposon_end(1)
                     /note="ApEinfo_label=transposon_end
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=#ff6666
                     ApEinfo_fwdcolor=#ff6666"
     misc_feature    1383
                     /label=transposon_end(1)
                     /note="ApEinfo_label=transposon_end
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=#ff6666
                     ApEinfo_fwdcolor=#ff6666"
     protein_bind    1392..1623
                     /gene="mutant version of attP"
                     /label=attP1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     CDS             complement(2019..2324)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(2669..3328)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3329..3431)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3576..3807)
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     misc_feature    complement(3816)
                     /label=transposon_end
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=#ff6666
                     ApEinfo_fwdcolor=#ff6666"
     misc_feature    complement(3817..3834)
                     /label=transposon_end
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=#ff6666
                     ApEinfo_fwdcolor=#ff6666"
     misc_feature    complement(3835..3849)
                     /label=nexteraB
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=#ff8000
                     ApEinfo_fwdcolor=#ff8000"
     misc_feature    join(3856..3875,3877..3905,3907..4055)
                     /label=polyA_rbcs
                     /note="ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 
                     0} width 5 offset 0
                     ApEinfo_revcolor=#008040
                     ApEinfo_fwdcolor=#008040"
     misc_feature    4068..4115
                     /label=LacZa-2
     misc_feature    4072..6332
                     /label=PMD19-2
     protein_bind    4135..4151
                     /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(4159..4189)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4204..4225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4513..5101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5272..6132)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6133..6237)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt agaactcggt acgcgcggat
      421 cttccagaga ttaaagaatt cgcccttata tattgatatt tatatcattt tacgtttctc
      481 gttcagcttt cttgtacaaa gtggtctcga ggatgcaaga cccttcctct atataaggaa
      541 gttcatttca tttggagagg tctagaatga gatctatgag taaaggagaa gaacttttca
      601 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
      661 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
      721 ctactggaaa actacctgtt ccatggccaa ccctggtcac caccctgacc tacggcgtgc
      781 agtgcttctc ccgttaccct gatcatatga agcggcacga cttcttcaag agcgccatgc
      841 ctgagggata cgtgcaggag aggaccatct tcttcaagga cgacgggaac tacaagacac
      901 gtgctgaagt caagtttgag ggagacaccc tcgtcaacag gatcgagctt aagggaatcg
      961 atttcaagga ggacggaaac atcctcggcc acaagttgga atacaactac aactcccaca
     1021 acgtatacat catggccgac aagcaaaaga acggcatcaa agccaacttc aagacccgcc
     1081 acaacatcga agacggcggc gtgcaactcg ctgatcatta tcaacaaaat actccaattg
     1141 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     1201 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     1261 ttacacatgg catggtagaa ctatacaaac accaccacca ccaccactaa gatatcgccc
     1321 agaattaatt ccccaactgc acagaacaaa tcgtcggcag cgtcagatgt gtataagaga
     1381 cagagctcat caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat
     1441 tgatgagcaa tgctttttta taatgccaac tttgtacaaa aaagctgaac gagaaacgta
     1501 aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata
     1561 ctgtaaaaca caacatatcc agtcactatg aatcaactac ttagatggta ttagtgacct
     1621 gtagtcgacc gacagccttc caaatgttct tcgggtgatg ctgccaactt agtcgaccga
     1681 cagccttcca aatgttcttc tcaaacggaa tcgtcgtatc cagcctactc gctattgtcc
     1741 tcaatgccgt attaaatcat aaaaagaaat aagaaaaaga ggtgcgagcc tcttttttgt
     1801 gtgacaaaat aaaaacatct acctattcat atacgctagt gtcatagtcc tgaaaatcat
     1861 ctgcatcaag aacaatttca caactcttat acttttctct tacaagtcgt tcggcttcat
     1921 ctggattttc agcctctata cttactaaac gtgataaagt ttctgtaatt tctactgtat
     1981 cgacctgcag actggctgtg tataagggag cctgacattt atattcccca gaacatcagg
     2041 ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc ttccccgata
     2101 acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc atccccgata
     2161 tgcaccaccg ggtaaagttc acgggagact ttatctgaca gcagacgtgc actggccagg
     2221 gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc cacaaacaga
     2281 cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc agcccctgtt
     2341 ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg acctcagcca tcccttcctg
     2401 attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca tggttgtgct
     2461 taccagaccg gagatattga catcatatat gccttgagca actgatagct gtcgctgtca
     2521 actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt cgggtataca
     2581 tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac tgttatctgg
     2641 cttttagtaa gccggatcca cgcggcgttt acgccccgcc ctgccactca tcgcagtact
     2701 gttgtaattc attaagcatt ctgccgacat ggaagccatc acagacggca tgatgaacct
     2761 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
     2821 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
     2881 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
     2941 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
     3001 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
     3061 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg aactccggat
     3121 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     3181 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     3241 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     3301 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     3361 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     3421 cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag
     3481 ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc
     3541 aaagtgcgtc gggtgatgct gccaacttag tcgactacag gtcactaata ccatctaagt
     3601 agttgattca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat
     3661 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct
     3721 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     3781 ctatcagtca aaataaaatc attatttgat ggtacctgtc tcttatacac atctccgagc
     3841 ccacgagacc ccgggtttcc ctttgctttt gtgtagacct caaaacttta tcccccatct
     3901 ttgatcttat cccttgtttt tctgcttttt tcttctttct tgggttttaa tttccggact
     3961 taacgtttgt tttccggttt gcgagacata ttctatcgga ttctcaactg tctgatgaaa
     4021 taaatatgta atgttctata agtctttcaa tttgatggga taagcttgtt aatcgtcgaa
     4081 cggcaggcgt gcaaacttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4141 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4201 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4261 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4321 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4381 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4441 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4501 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4561 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4621 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4681 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4741 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4801 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4861 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4921 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4981 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5041 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5101 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5161 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5221 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5281 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5341 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5401 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5461 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5521 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5581 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5641 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5701 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5761 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5821 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5881 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5941 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6001 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6061 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6121 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6181 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatggggg ttccgcgcac
     6241 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6301 taaaaatagg cgtatcacga ggccctttcg tc
//
