LOCUS       Exported                5077 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of LC3B.
ACCESSION   .
VERSION     .
KEYWORDS    HA-MAP1LC3B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5077)
  AUTHORS   Stefely JA, Zhang Y, Freiberger EC, Kwiecien NW, Thomas HE, Davis 
            AM, Lowry ND, Vincent CE, Shishkova E, Clark NA, Medvedovic M, Coon 
            JJ, Pagliarini DJ, Mercer CA
  TITLE     Mass spectrometry proteomics reveals a function for mammalian 
            CALCOCO1 in MTOR-regulated selective autophagy.
  JOURNAL   Autophagy. 2020 Jan 23. doi: 10.1080/15548627.2020.1719746.
  PUBMED    31971854
REFERENCE   2  (bases 1 to 5077)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5077
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             907..933
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1324..1353
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1364..1373
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    1377..1511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1414..1433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1468..1487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1580..1937
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(1603..1623)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1788..1923
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1850..1869
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(1957..1974)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1957..1973)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1966..1988)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2186..2641
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2273..2292)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2483..2504
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2658..2677)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2777..2799
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2837..2855)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2923..3027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3028..3888
                     /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"
     primer_bind     complement(3246..3265)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4059..4647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4548..4567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4801..4818
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4935..4956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4971..5001
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5009..5025
                     /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)."
     primer_bind     5014..5036
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5033..5049
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5033..5049
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgtacc tcgcgagcca
      901 gccatgtacc catacgacgt cccagactac gctggcggcc gcccgtcgga gaagaccttc
      961 aagcagcgcc gcaccttcga acaaagagta gaagatgtcc gacttattcg agagcagcat
     1021 ccaaccaaaa tcccggtgat aatagaacga tacaagggtg agaagcagct tcctgttctg
     1081 gataaaacaa agttccttgt acctgaccat gtcaacatga gtgagctcat caagataatt
     1141 agaaggcgct tacagctcaa tgctaatcag gccttcttcc tgttggtgaa cggacacagc
     1201 atggtcagcg tctccacacc aatctcagag gtgtatgaga gtgagaaaga tgaagatgga
     1261 ttcctgtaca tggtctatgc ctcccaagaa acatttggaa tgaagctaag tgtatctaga
     1321 atggagcaga agctgatctc cgaggaggac ctgtgaaagc ttggccgcca tggcccaact
     1381 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1441 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1501 atgtctggat cgggaattaa ttcggcgcag caccatggcc tgaaataacc tctgaaagag
     1561 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     1621 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1681 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1741 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     1801 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     1861 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     1921 gcctaggctt ttgcaaaaag ctgttaacag cttggcactg gccgtcgttt tacaacgtcg
     1981 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     2041 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     2101 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2161 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2221 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2281 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2341 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2401 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2461 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2521 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2581 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     2641 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     2701 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2761 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     2821 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     2881 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2941 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3001 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3061 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3121 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3181 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3241 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3301 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3361 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3421 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3481 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3541 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3601 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3661 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3721 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3781 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3841 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3901 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3961 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4021 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4081 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4141 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4201 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4261 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4321 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4381 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4441 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4501 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4561 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4621 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4681 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4741 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4801 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4861 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     4921 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     4981 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     5041 agctatgaca tgattacgaa ttaattcgag ctcgccc
//
