LOCUS       Exported                8315 bp ds-DNA     circular SYN 25-DEC-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8315)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Dec 25, 2020 from SnapGene 5.1.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8315
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..2848
                     /label=Backbone
                     /note="color: #ffcc99"
     protein_bind    11..27
                     /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"
     promoter        complement(35..65)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 35 .. 41 / #ffffff / -10
                        2: 42 .. 59 / #ffffff
                        3: 60 .. 65 / #ffffff / -35"
     protein_bind    80..101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(389..974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(1145..2005)
                     /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: 1145 .. 1936 / #ccffcc
                        2: 1937 .. 2005 / #ccffcc / signal sequence
                     Cleavage site after base 1936"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2006..2110)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      2137..2592
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     2732..2749
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    2852..3551
                     /label=5' homology arm
                     /note="color: #3366ff"
     misc_feature    2864..2883
                     /label=Primer 610
                     /note="color: #a6acb3; direction: RIGHT"
     promoter        3565..4743
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    complement(3677..3697)
                     /label=Primer 764
                     /note="color: #a6acb3; direction: LEFT"
     intron          3796..4734
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
                     /note="color: #------"
     misc_feature    4757..4762
                     /label=Kozak
                     /note="color: #ff6600"
     CDS             4763..5473
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    5591..5712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     protein_bind    5753..5786
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     promoter        5823..6319
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    complement(5911..5930)
                     /label=Primer 665
                     /note="color: #a6acb3; direction: LEFT"
     CDS             6359..6958
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    7036..7488
                     /label=PGK polyA
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    7404..7425
                     /label=Primer 666
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    7539..7572
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    7608..8310
                     /label=3' homology arm
                     /note="color: #3366ff"
     misc_feature    complement(7964..7983)
                     /label=Primer 611
                     /note="color: #a6acb3; direction: LEFT"
ORIGIN
        1 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
       61 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      121 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      181 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      241 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      301 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
      841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      901 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      961 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1021 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1081 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1141 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1201 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1261 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1321 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1381 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1441 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1501 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1561 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1621 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1681 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1741 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1801 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1861 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1921 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1981 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2041 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2101 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     2161 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2221 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2281 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2341 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2401 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2461 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2521 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2581 cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     2641 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     2701 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctgat
     2761 ctatacattg aatcaatatt ggcaattagc catattagtc attggttata tagcataaat
     2821 caatattggc tattggccat tgcatacgtt tcttgtgaac tcatctcgtg cctgcaagac
     2881 atcaaaagtt tctccaaaga catcccttcc tctctgccct catcctatat caaggtctta
     2941 gctccttgaa gactgcatta atgtgtcttt tgccttcatc ccccccccac cctcccgctt
     3001 cctctagtcc ttgtgtttac caaaatactt ttgctaaaac ctgtatgtct agcttctgct
     3061 gatactctta gcaatactcc cttatttcct tctggcgcca tttgccaatc accagctaat
     3121 ggctttgctc tttgcatggt acctgtttct gtcctactag cttctagtct gaaaaggaag
     3181 cctttactag cagcttcaac ccagtcctcc ttaccttgca cctattaacg ttggttcaac
     3241 atgaagggga agtaagataa gtggaagagg atacacagga aaggtggaac tggattcagg
     3301 tgtctgtgtt ctgccattga cacttgggac tgcgttccag cttccacctg tggaaaccag
     3361 aaagcatttg ccaaaaattc taaacagagg gtctatttct gaagctgagg aatcacatgg
     3421 agtgaatagc atgggggaag gggattacta atgaggctga aagaaaagta ccaggaagag
     3481 actgacttca tgcataatcc ataaagccag gttttcttga aaggtgacct gctttcttgt
     3541 attcctgcaa gaaacggctc gaggggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     3601 cccacagtcc ccgagaagtt gtggggaggg gtcggcaatt gaagcggtgc ctagagaagg
     3661 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     3721 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     3781 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
     3841 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
     3901 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
     3961 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     4021 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     4081 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac
     4141 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
     4201 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
     4261 gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
     4321 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
     4381 gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
     4441 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
     4501 cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt
     4561 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
     4621 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
     4681 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
     4741 tgaggggatc gtcgacgcca ccatggtgag caagggcgag gaggataaca tggccatcat
     4801 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     4861 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     4921 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     4981 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     5041 cgagggcttc aattgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     5101 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     5161 cttcccctcc gacggccccg taatgcagtg tcgtaccatg ggctgggagg cctccactga
     5221 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     5281 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     5341 gctgcccggc gcctacaacg tcgacatcaa gttggacatc ctttcccaca acgaggacta
     5401 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     5461 gctgtacaag taatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     5521 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     5581 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5641 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5701 caatgtatct taacgcgatc gccagtacta gtgaacctct tcgagggacc taataacttc
     5761 gtatagcata cattatacga agttatatta agggttccgg atctcgacct cgaaattcta
     5821 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     5881 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagc
     5941 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     6001 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     6061 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     6121 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagcag ctgggaaggg
     6181 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     6241 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     6301 ctcatctccg ggcctttcga cctgcatcca tctagatctc gagcagctga agcttaccat
     6361 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     6421 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     6481 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     6541 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     6601 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     6661 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     6721 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     6781 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     6841 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     6901 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
     6961 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgatgat
     7021 atcagatccc cgggatgcag aaattgatga tctattaaac aataaagatg tccactaaaa
     7081 tggaagtttt tcctgtcata ctttgttaag aagggtgaga acagagtacc tacattttga
     7141 atggaaggat tggagctacg ggggtggggg tggggtggga ttagataaat gcctgctctt
     7201 tactgaaggc tctttactat tgctttatga taatgtttca tagttggata tcataattta
     7261 aacaagcaaa accaaattaa gggccagctc attcctccca ctcatgatct atagatctat
     7321 agatctctcg tgggatcatt gtttttctct tgattcccac tttgtggttc taagtactgt
     7381 ggtttccaaa tgtgtcagtt tcatagcctg aagaacgaga tcagcagcct ctgttccaca
     7441 tacacttcat tctcagtatt gttttgccaa gttctaattc catcagaagc tggtcgagat
     7501 cctaagcttg gctggacgta aactcctctt cagacctaat aacttcgtat agcatacatt
     7561 atacgaagtt atattaaggg ttattgaata tgatcggaat tgcggccctg acctgttggg
     7621 gtctttaaag ctcaaggaaa aaggccatag ttgatttctc ctaaatcaag atagagtcca
     7681 attaactttt tttttttttt aagatggagt ctcactctgt cgccaggctg gactgcagtg
     7741 gcgtgatctc agctcactgc aacctctgcc tcccaggttc aggcgattct tctgtctcag
     7801 cctcccgagt agctgggact acaagtgtgg gccaccatgc ccagctagtt ttgtattttt
     7861 agtagagatg ggatttcacc atcttggcta ggatggtctt gatctcttga ccttgtgatt
     7921 cgcctgcctc tgcctcccag agtgctgaga ttacaggcgt gagccacagt gcccaatttc
     7981 catgggtttt caagaaaaac ttaacttact taaacttcag atcacttcct taaaggaaca
     8041 tactgagcat agcttggctg gttagaaatt aggaagacta gcttgggcta catggtaaga
     8101 ccctgcctct acaaaaaata agaaaaaagt tagtggcatg tgcctgtagt tccatctact
     8161 tgggaggctg aggtgagagg atcgcttgag cccaggaggt tgaagctgca gtgagccacg
     8221 attgcaccac tgtactccag cctgggtgac acagagtgag accctgtctc caaacaaaat
     8281 taggaagggt ttcagagagg aaataaacac ggccg
//
