LOCUS       Exported                5564 bp DNA     circular SYN 05-JUL-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5564)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 11, 2022 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5564
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
                     /note="color: black; sequence: CGCAAATGGGCGGTAGGCGTG"
     primer_bind     complement(631..650)
                     /label=Vector.REV
                     /note="color: black; sequence: ccgatttccatggcggtctc"
     primer_bind     636..656
                     /label=Fragment.FOR
                     /note="color: black; sequence: cgccatggaaatcggtactgg"
     CDS             643..1527
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /note="color: #cc99b2"
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(1061..1079)
                     /label=Halo-mGold-F152A_rev
                     /note="color: black; sequence: GTCTCGCGGGCAAATTCTG"
     primer_bind     1080..1101
                     /label=Halo-mGold-F152A_for
                     /note="color: black; sequence: CTTCCAGGCCgcCCGCACCACC"
     primer_bind     complement(1108..1125)
                     /label=Halo-mGold-V167G_rev
                     /note="color: black; sequence: GATGATCAGCTTGCGGCC"
     primer_bind     1126..1153
                     /label=Halo-mGold-V167G_for
                     /note="color: black; sequence: 
                     GATCAGAACGgcTTTATCGAGGGTACGC"
     primer_bind     complement(1516..1534)
                     /label=FRAGMENT_rev
                     /note="color: black; sequence: tgctcacgccggaaatctc"
     primer_bind     1519..1539
                     /label=Vector.FOR
                     /note="color: black; sequence: atttccggcgtgagcaagggc"
     CDS             1528..2244
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /note="This feature has 2 segments:
                        1: 1528 .. 1530 / #e1ff2f / 1a
                        2: 1531 .. 2244 / #e1ff2f"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     regulatory      1528
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     primer_bind     complement(1570..1591)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     complement(1831..1850)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: black; sequence: GTCTTGTAGTTGCCGTCGTC"
     primer_bind     2178..2199
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     polyA_signal    2367..2488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(2404..2423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: black; sequence: GAAATTTGTGATGCTATTGC"
     primer_bind     2458..2477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: black; sequence: GTGGTTTGTCCAAACTCATC"
     rep_origin      complement(2495..2950)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(2632..2653)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: black; sequence: GTGGACTCTTGTTCCAAACTGG"
     primer_bind     2844..2863
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: black; sequence: AGGGAAGAAAGCGAAAGGAG"
     promoter        2977..3081
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        3083..3440
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(3106..3126)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
                     /note="color: black; sequence: ACCCTAACTGACACACATTCC"
     rep_origin      3291..3426
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     3353..3372
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     primer_bind     complement(3376..3387)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
                     /note="color: black; sequence: TATTTATGCAGAGGCCGAGG"
     CDS             3475..4269
                     /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     complement(3529..3548)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
                     /note="color: black; sequence: GCCCAGTCATAGCCGAATAG"
     primer_bind     4139..4158
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
                     /note="color: black; sequence: CGTTGGCTACCCGTGATATT"
     primer_bind     complement(4457..4476)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
                     /note="color: black; sequence: TTGTCTCCTTCCGTGTTTCA"
     polyA_signal    4501..4548
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     rep_origin      4877..5465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     5366..5385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
ORIGIN
        1 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatccgctag cgctaccgga ctcagatctc gagaccgcca tggaaatcgg tactggcttt
      661 ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt
      721 ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg
      781 cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga cctgatcggt
      841 atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt ccgcttcatg
      901 gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca cgactggggc
      961 tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg tattgcattt
     1021 atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc ccgcgagacc
     1081 ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca gaacgttttt
     1141 atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat
     1201 taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt cccaaacgag
     1261 ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata catggactgg
     1321 ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt tctgatccca
     1381 ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc
     1441 ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga gatcgcgcgc
     1501 tggctgtcca cgctcgagat ttccggcgtg agcaagggcg aggagctgtt caccggggtg
     1561 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1621 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1681 aagctgcccg tgccctggcc caccctcgtg accagcctgg gctacggcct gcagtgcttc
     1741 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1801 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1861 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1921 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1981 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     2041 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2101 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     2161 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2221 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     2281 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2341 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2401 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2461 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     2521 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2581 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2641 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2701 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2761 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2821 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2881 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2941 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3001 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3061 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3121 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3181 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3241 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3301 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3361 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3421 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3481 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3541 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3601 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3661 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3721 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3781 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3841 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3901 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3961 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4021 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4081 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4141 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4201 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4261 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4321 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4381 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4441 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4501 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4561 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4621 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4681 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4741 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4801 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4861 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4921 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4981 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5041 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5101 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5161 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5221 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5281 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5341 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5401 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5461 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5521 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgt
//
