LOCUS       Exported                5053 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of GABARAPL1.
ACCESSION   .
VERSION     .
KEYWORDS    HA-GABARAPL1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5053)
  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 5053)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5053
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             920..946
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1313..1342
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1353..1362
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    1366..1500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1403..1422)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1457..1476
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1569..1926
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(1592..1612)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1777..1912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1839..1858
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(1946..1963)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1946..1962)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1955..1977)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2175..2630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2262..2281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2472..2493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2647..2666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2766..2788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2826..2844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2912..3016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3017..3877
                     /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(3235..3254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4048..4636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4537..4556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4790..4807
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4924..4945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4960..4990
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4998..5014
                     /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     5003..5025
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5022..5038
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5022..5038
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgt
      901 acctcgcgag ccagccatgt acccatacga cgtcccagac tacgctggcg gccgcaagtt
      961 ccagtacaag gaggaccatc cctttgagta tcggaaaaag gaaggagaaa agatccggaa
     1021 gaaatatccg gacagggtcc ccgtgattgt agagaaggct ccaaaagcca gggtgcctga
     1081 tctggacaag aggaagtacc tagtgccctc tgaccttact gttggccagt tctacttctt
     1141 aatccggaag agaatccacc tgagacctga ggacgcctta ttcttctttg tcaacaacac
     1201 catccctccc accagtgcta ccatgggcca actgtatgag gacaatcatg aggaagacta
     1261 ttttctgtat gtggcctaca gtgatgagag tgtctatggg aaatctagaa tggagcagaa
     1321 gctgatctcc gaggaggacc tgtgaaagct tggccgccat ggcccaactt gtttattgca
     1381 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1441 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     1501 gggaattaat tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     1561 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     1621 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1681 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1741 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     1801 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     1861 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     1921 tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     1981 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     2041 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2101 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     2161 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2221 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2281 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2341 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     2401 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2461 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     2521 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2581 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     2641 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     2701 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2761 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2821 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2881 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2941 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3001 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3061 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3121 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3181 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3241 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3301 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3361 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3421 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3481 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3541 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3601 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3661 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3721 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3781 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3841 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3901 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3961 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4021 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4081 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4141 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4201 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4261 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4321 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4381 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4441 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4501 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4561 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4621 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4681 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4741 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4801 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     4861 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     4921 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     4981 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat
     5041 gattacgaat taa
//
