LOCUS       Exported                8826 bp ds-DNA     circular SYN 01-AUG-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8826)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Aug 1, 2019 from SnapGene 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8826
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    194..215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        230..260
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 230 .. 235 / #ffffff / -35
                        2: 236 .. 253 / #ffffff
                        3: 254 .. 260 / #ffffff / -10"
     protein_bind    268..284
                     /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)."
                     /note="color: #31849b"
     primer_bind     292..308
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     misc_binding    complement(474..1174)
                     /label=piggyBacL
                     /note="color: #f7a68a; direction: LEFT"
     terminator      complement(1214..1447)
                     /label=SV40
                     /note="color: #000000; direction: LEFT"
     primer_bind     complement(1320..1334)
                     /label=Primer 21
                     /note="color: black; sequence: TCGACCTCGAAACTTGTTTATTGCA"
     misc_feature    complement(1455..2135)
                     /label=Dsred
                     /note="Dsred"
                     /note="color: #ff0000; direction: LEFT"
     primer_bind     2142..2165
                     /label=Primer 9
                     /note="color: black; sequence: GTCACTTGGTTGTTCACGATCTTG"
     promoter        complement(2146..2737)
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
                     /note="color: #ffffff; direction: LEFT"
     enhancer        2741..3223
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
                     /note="color: #a6acb3"
     primer_bind     complement(3196..3223)
                     /label=Primer 2
                     /note="color: black; sequence: 
                     CGCGTAAAACACAATCAAGTATGAGTCA"
     misc_feature    3231..3777
                     /label=Opie2 promoter
                     /note="Opie2 promoter"
                     /note="color: #ff8000; direction: RIGHT"
     primer_bind     3237..3257
                     /label=Primer 16
                     /note="color: black; sequence: CATACTCGGTGGCCTCCCCAC"
     primer_bind     complement(3759..3777)
                     /label=Primer 20
                     /note="color: black; sequence: CACCAGAGACAGGTTGCGG"
     CDS             3786..4580
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     3786..3810
                     /label=Primer 27
                     /note="color: black; sequence: ATGATTGAACAAGATGGATTGCACG"
     primer_bind     complement(4557..4580)
                     /label=Primer 30
                     /note="color: black; sequence: TCAGAAGAACTCGTCAAGAAGGCG"
     primer_bind     complement(4560..4580)
                     /label=Primer 29
                     /note="color: black; sequence: TCAGAAGAACTCGTCAAGAAG"
     primer_bind     4587..4611
                     /label=Primer 21
                     /note="color: black; sequence: TCGACCTCGAAACTTGTTTATTGCA"
     polyA_signal    4597..4731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(4725..4749)
                     /label=Primer 18
                     /note="color: black; sequence: CGGGCCCCCCCTCGACATGATCCAG"
     primer_bind     complement(4735..4749)
                     /label=Primer 17
                     /note="color: black; sequence: CGGGCCCCCCCTCGA"
     misc_binding    4803..5800
                     /label=piggyBacR
                     /note="color: #f7a68a"
     primer_bind     complement(6518..6534)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     promoter        7008..7112
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             7113..7973
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 7113 .. 7181 / #ccffcc / signal sequence
                        2: 7182 .. 7973 / #ccffcc
                     Cleavage site after base 7181"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8144..8732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
       61 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
      121 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      181 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      241 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      301 gctatgacca tgattacgaa ttcgagctcg gtacccgggg atcctctaga gtcgacgctc
      361 gcgcgacttg gtttgccatt ctttagcgcg cgtcgcgtca cacagcttgg ccacaatgtg
      421 gtttttgtca aacgaagatt ctatgacgtg tttaaagttt aggtcgagta aagcgcaaat
      481 cttttttaac cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg
      541 ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc
      601 ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa
      661 cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc
      721 atacgataat tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat
      781 tttcttgtta tagatatcgt gactaatata taataaaatg ggtagttctt tagacgatga
      841 gcatatcctc tctgctcttc tgcaaagcga tgacgagctt gttggtgagg attctgacag
      901 tgaaatatca gatcacgtaa gtgaagatga cgtccagagc gatacagaag aagcgtttat
      961 agatgaggta catgaagtgc agccaacgtc aagcggtagt gaaatattag acgaacaaaa
     1021 tgttattgaa caaccaggtt cttcattggc ttctaacaga atcttgacct tgccacagag
     1081 gactattaga ggtaagaata aacattgttg gtcaacttca aagtccacga ggcgtagccg
     1141 agtctctgca ctgaacattg tcagatccga gatcggccgg cctaggcgcg ccgtacgcgt
     1201 atcgataagc tttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1261 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1321 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1381 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     1441 tagagtcgcg gccgctacag gaacaggtgg tggcggccct cggtgcgctc gtactgctcc
     1501 acgatggtgt agtcctcgtt gtgggaggtg atgtccagct tggagtccac gtagtagtag
     1561 ccgggcagct gcacgggctt cttggccatg tagatggact tgaactccac caggtagtgg
     1621 ccgccgtcct tcagcttcag ggccttgtgg atctcgccct tcagcacgcc gtcgcggggg
     1681 tacaggcgct cggtggaggc ctcccagccc atggtcttct tctgcattac ggggccgtcg
     1741 gaggggaagt tcacgccgat gaacttcacc ttgtagatga agcagccgtc ctgcagggag
     1801 gagtcctggg tcacggtcac cacgccgccg tcctcgaagt tcatcacgcg ctcccacttg
     1861 aagccctcgg ggaaggacag cttcttgtag tcggggatgt cggcggggtg cttcacgtac
     1921 accttggagc cgtactggaa ctggggggac aggatgtccc aggcgaaggg cagggggccg
     1981 cccttggtca ccttcagctt cacggtgttg tggccctcgt aggggcggcc ctcgccctcg
     2041 ccctcgatct cgaactcgtg gccgttcacg gtgccctcca tgcgcacctt gaagcgcatg
     2101 aactccttga tgacgttctt ggaggagcgc accatctcga ggtcacttgg ttgttcacga
     2161 tcttgtcgcc gccagtgtca acttgcaact gaaacaatat ccaacatgaa cgtcaattta
     2221 tactgcccta atggcgaaca cgataacaat atttctttta ttatgccctc taaaaccaac
     2281 gcggttatcg tttatttatt caaattagat atagaacatc cgccgacata caatgttaat
     2341 gcaaaaacgc gtttggtgag cggatacgaa aacagtcggc cgataaacat taatctgagg
     2401 tcgataacac cgtccttgaa cggaacacga ggagcgtacg tgatcagctg cattcgcgcg
     2461 ccgcgccttt atcgagattt atttgcatac aacaagtaca ctgcgccgtt gggatttgtg
     2521 gtaacgcgca cacatgcaga gctgcaagtg tggcacattt tgtctgtgcg caaaaccttt
     2581 gaagccaaaa gtacgaggtc cgttacgggc atgctagcgc acacggacaa tggacccgac
     2641 aaattctacg ccaaggattt aatgataatg tcgggcaacg tatccgttca ttttatcaat
     2701 aacctacaaa aatgtcgcgc gcatcacaaa gacatcgacg cgcgtagaat tctacccgta
     2761 aagcgagttt agttatgagc catgtgcaaa acatgacatc agcttttatt tttataacaa
     2821 atgacatcat ttcttgattg tgttttacac gtagaattct actcgtaaag cgagttcagt
     2881 tttgaaaaac aaatgacatc atctttttga ttgtgcttta caagtagaat tctacccgta
     2941 aatcaagttc ggttttgaaa aacaaatgag tcatattgta tgatatcata ttgcaaaaca
     3001 aatgactcat caatcgatcg tgcgttacac gtagaattct actcgtaaag cgagtttatg
     3061 agccgtgtgc aaaacatgac atcatctcga tttgaaaaac aaatgacatc atccactgat
     3121 cgtgcgttac aagtagaatt ctactcgtaa agccagttcg gttatgagcc gtgtgcaaaa
     3181 catgacatca gcttatgact catacttgat tgtgttttac gcggtttaaa cgcgcccata
     3241 ctcggtggcc tccccaccac caactttttt gcactgcaaa aaaacacgct tttgcacgcg
     3301 ggcccataca tagtacaaac tctacgtttc gtagactatt ttacataaat agtctacacc
     3361 gttgtatacg ctccaaatac actaccacac attgaacctt tttgcagtgc aaaaaagtac
     3421 gtgtcggcag tcacgtaggc cggccttatc gggtcgcgtc ctgtcacgta cgaatcacat
     3481 tatcggaccg gacgagtgtt gtcttatcgt gacaggacgc cagcttcctg tgttgctaac
     3541 cgcagccgga cgcaactcct tatcggaaca ggacgcgcct ccatatcagc cgcgcgttat
     3601 ctcatgcgcg tgaccggaca cgaggcgccc gtcccgctta tcgcgcctat aaatacagcc
     3661 cgcaacgatc tggtaaacac agttgaacag catctgttac agcgacacaa catgagccgg
     3721 tccaacaacg ccaacgcgcc cacgccatcc aaccgccgcc gcaacctgtc tctggtgtta
     3781 attaaatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3841 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3901 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3961 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4021 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4081 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4141 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4201 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4261 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4321 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4381 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4441 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4501 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4561 ttcttgacga gttcttctga agatcttcga cctcgaaact tgtttattgc agcttataat
     4621 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4681 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat catgtcgagg
     4741 gggggcccga tttaaataaa ccccgggcga gctcgaatta accattgtgg gaaccgtgcg
     4801 atcaaacaaa cgcgagatac cggaagtact gaaaaacagt cgctccaggc cagtgggaac
     4861 atcgatgttt tgttttgacg gaccccttac tctcgtctca tataaaccga agccagctaa
     4921 gatggtatac ttattatcat cttgtgatga ggatgcttct atcaacgaaa gtaccggtaa
     4981 accgcaaatg gttatgtatt ataatcaaac taaaggcgga gtggacacgc tagaccaaat
     5041 gtgttctgtg atgacctgca gtaggaagac gaataggtgg cctatggcat tattgtacgg
     5101 aatgataaac attgcctgca taaattcttt tattatatac agccataatg tcagtagcaa
     5161 gggagaaaag gttcaaagtc gcaaaaaatt tatgagaaac ctttacatga gcctgacgtc
     5221 atcgtttatg cgtaagcgtt tagaagctcc tactttgaag agatatttgc gcgataatat
     5281 ctctaatatt ttgccaaatg aagtgcctgg tacatcagat gacagtactg aagagccagt
     5341 aatgaaaaaa cgtacttact gtacttactg cccctctaaa ataaggcgaa aggcaaatgc
     5401 atcgtgcaaa aaatgcaaaa aagttatttg tcgagagcat aatattgata tgtgccaaag
     5461 ttgtttctga ctgactaata agtataattt gtttctatta tgtataagtt aagctaatta
     5521 cttattttat aatacaacat gactgttttt aaagtacaaa ataagtttat ttttgtaaaa
     5581 gagagaatgt ttaaaagttt tgttacttta tagaagaaat tttgagtttt tgtttttttt
     5641 taataaataa ataaacataa ataaattgtt tgttgaattt attattagta tgtaagtgta
     5701 aatataataa aacttaatat ctattcaaat taataaataa acctcgatat acagaccgat
     5761 aaaacacatg cgtcaatttt acgcatgatt atctttaacg tacgtcacaa tatgattatc
     5821 tttctagggt taaataatag tttctaattt ttttattatt cagcctgctg tcgtgaatac
     5881 cgtatatctc aacgctgtct gtgagattgt cgtattctag cctttttagt ttttcgctca
     5941 tcgacttgat attgtccgac acattttcgt cgatttgcgt tttgatcaaa gacttgagca
     6001 gagacacgtt aatcaactgt tcaaattgat ccatattaac gatatcaacc cgatgcgtat
     6061 atggtgcgta aaatatattt tttaaccctc ttatactttg cactctgcgt taatacgcgt
     6121 tcgtgtacag acgtaatcat gttttctttt ttggataaaa ctcctactga gtttgacctc
     6181 atattagacc ctcacaagtt gcaaaacgtg gcatttttta ccaatgaaga atttaaagtt
     6241 attttaaaaa atttcatcac agatttaaag aagaaccaaa aattaaatta tttaatcgac
     6301 cagttaatca acgtgtacac agagcgcaaa aaacacgcag cccgacgtgt tggctaaaat
     6361 tattaaatca acttgtgtta tagtcacgat ttgccgtcca acgtgttcct caaaaagttg
     6421 aagaccaaca agtttacgga cactagttaa ttatttgatt ttgccccact tcattttgtg
     6481 ggatcacaat tttgttatat tttaaacaaa gcttggcact ggccgtcgtt ttacaacgtc
     6541 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6601 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6661 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6721 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     6781 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     6841 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     6901 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     6961 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     7021 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     7081 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     7141 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     7201 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     7261 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     7321 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     7381 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7441 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7501 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7561 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7621 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7681 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7741 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7801 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7861 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7921 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     7981 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     8041 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     8101 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     8161 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8221 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8281 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8341 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8401 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8461 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8521 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8581 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8641 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     8701 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     8761 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8821 tggata
//
