LOCUS       Exported                7763 bp ds-DNA     circular SYN 04-MAY-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7763)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported May 4, 2021 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7763
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        386..661
                     /label=chicken beta-actin promoter
                     /note="color: #ffffff; direction: RIGHT"
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
                     /note="color: #------"
     CDS             1762..2736
                     /codon_start=1
                     /label=Chronos
                     /note="color: #a6acb3"
                     /translation="METAATMTHAFISAVPSAEATIRGLLSAAAVVTPAADAHGETSNA
                     TTAGADHGCFPHINHGTELQHKIAVGLQWFTVIVAIVQLIFYGWHSFKATTGWEEVYVC
                     VIELVKCFIELFHEVDSPATVYQTNGGAVIWLRYSDWLLTCPVILIHLSNLTGLHEEYS
                     KRTMTILVTDIGNIVWGITAAFTKGPLKILFFMIGLFYGVTCFFQIAKVYIESYHTLPK
                     GVCRKICKIMAYVFFCSWLMFPVMFIAGHEGLGLITPYTSGIGHLILDLISKNTWGFLG
                     HHLRVKIHEHILIHGDIRKTTTINVAGENMEIETFVDEEEEGGV"
     misc_feature    1990..2007
                     /label=seq primer 3
                     /note="color: #a6acb3"
     misc_feature    2737..2751
                     /label=Linker
                     /note="color: #a6acb3"
     CDS             2752..3462
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /note="color: #ff0000"
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    3463..3471
                     /label=Linker
                     /note="color: #a6acb3"
     misc_feature    3472..3669
                     /label=Soma-targeting motif
                     /note="color: #339966"
     intron          3686..3819
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
                     /note="color: #------"
     misc_feature    3828..4416
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     misc_feature    3854..3873
                     /label=seq primer 5
                     /note="color: #a6acb3"
     CDS             complement(4299..4310)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 4298"
                     /translation="IEGR"
     polyA_signal    4449..4560
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     polyA_site      4852..4907
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5266..5282)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5290..5306
                     /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(5314..5344)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5314 .. 5320 / #ffffff / -10
                        2: 5321 .. 5338 / #ffffff
                        3: 5339 .. 5344 / #ffffff / -35"
     protein_bind    5359..5380
                     /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        5439..5634
                     /label=SV40 promoter
                     /note="SV40 early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      5485..5620
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     polyA_signal    5640..5774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(6013..6601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6772..7632)
                     /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: 6772 .. 7563 / #ccffcc
                        2: 7564 .. 7632 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    6813..6832
                     /label=Seq primer 8
                     /note="color: #a6acb3"
     misc_feature    7460..7479
                     /label=Seq primer 7
                     /note="color: #a6acb3"
     promoter        complement(7633..7737)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgg taccgcgggc
     1741 ccgggatcca ccggtgctag tatggaaaca gccgccacaa tgacccacgc ctttatctca
     1801 gccgtgccta gcgccgaagc cacaattaga ggcctgctga gcgccgcagc agtggtgaca
     1861 ccagcagcag acgctcacgg agaaacctct aacgccacaa cagccggagc cgatcacggt
     1921 tgcttccccc acatcaacca cggaaccgag ctgcagcaca agatcgcagt gggactccag
     1981 tggttcaccg tgatcgtggc tatcgtgcag ctcatcttct acggttggca cagcttcaag
     2041 gccacaaccg gctgggagga ggtctacgtc tgcgtgatcg agctcgtcaa gtgcttcatc
     2101 gagctgttcc acgaggtcga cagcccagcc acagtgtacc agaccaacgg aggagccgtg
     2161 atttggctgc ggtacagcga ttggctcctg acttgccccg tgatcctgat ccacctgagc
     2221 aacctgaccg gactgcacga agagtacagc aagcggacca tgaccatcct ggtgaccgac
     2281 atcggcaaca tcgtgtgggg gatcacagcc gcctttacaa agggccccct gaagatcctg
     2341 ttcttcatga tcggcctgtt ctacggcgtg acttgcttct tccagatcgc caaggtgtat
     2401 atcgagagct accacaccct gcccaaaggc gtctgccgga agatttgcaa gatcatggcc
     2461 tacgtcttct tctgctcttg gctgatgttc cccgtgatgt tcatcgccgg acacgaggga
     2521 ctgggcctga tcacacctta caccagcgga atcggccacc tgatcctgga tctgatcagc
     2581 aagaacactt ggggcttcct gggccaccac ctgagagtga agatccacga gcacatcctg
     2641 atccacggcg acatccggaa gacaaccacc atcaacgtgg ccggcgagaa catggagatc
     2701 gagaccttcg tcgacgagga ggaggaggga ggagtggcgg ccgcaccggt aatggtgtct
     2761 aagggcgaag agctgatcaa ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc
     2821 aacggccacc aattcaaatg cacaggtgaa ggagaaggca atccgtacat gggaactcaa
     2881 accatgagga tcaaagtcat cgagggagga cccctgccat ttgcctttga cattcttgcc
     2941 acgtcgttca tgtatggcag ccgtactttt atcaagtacc cgaaaggcat tcctgatttc
     3001 tttaaacagt cctttcctga gggttttact tgggaaagag ttacgagata cgaagatggt
     3061 ggagtcgtca ccgtcatgca ggacaccagc cttgaggatg gctgtctcgt ttaccacgtc
     3121 caagtcagag gggtaaactt tccctccaat ggtcccgtga tgcagaagaa gaccaagggt
     3181 tgggagccta atacagagat gatgtatcca gcagatggtg gtctgagggg atacactcat
     3241 atggcactga aagttgatgg tggtggccat ctgtcttgct ctttcgtaac aacttacagg
     3301 tcaaaaaaga ccgtcgggaa catcaagatg cccggtatcc atgccgttga tcaccgcctg
     3361 gaaaggttag aggaaagtga caatgaaatg ttcgtagtac aacgcgaaca cgcagttgcc
     3421 aagttcgccg ggcttggtgg tgggatggac gagctgtaca aggccctaga gcagtcccag
     3481 cccatcctca acaccaagga gatggccccg cagagcaagc ctccagagga gctggagatg
     3541 agcagcatgc ccagccccgt ggcccctctg cccgcacgca cggagggcgt catcgacatg
     3601 cggagcatgt ccagcattga cagcttcatc agctgtgcca cggacttccc tgaagccacc
     3661 agattctaag gactcgaggc cgcaggtaag tatcaaggtt acaagacagg tttaaggaga
     3721 ccacatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta
     3781 ttggtcttac tgacatccac tttgcctttc tctccacagg tgtcgacaat caacctctgg
     3841 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3901 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3961 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     4021 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     4081 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     4141 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     4201 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     4261 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     4321 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     4381 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattcgagct cggtacgatc
     4441 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4501 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4561 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4621 ggaggattgg gaagacaata gcaggccagc ttttgttccc tttagggtta attgcgcgct
     4681 tggcgctaat catggtcata gctgtttcca ctcctcaggt gcaggctgcc tatcagaagg
     4741 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     4801 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     4861 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     4921 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     4981 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
     5041 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     5101 ttttatattt tgtttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     5161 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     5221 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     5281 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5341 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5401 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     5461 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5521 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5581 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     5641 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5701 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5761 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5821 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5881 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5941 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6001 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6061 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6121 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6181 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     6241 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6301 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6361 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6421 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6481 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6541 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6601 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6661 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6721 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6781 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6841 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6901 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6961 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7021 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7081 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7141 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7201 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7261 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7321 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7381 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7441 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7501 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7561 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7621 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7681 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7741 atttccccga aaagtgccac ctg
//
