LOCUS       pUC57-gCarvi-T2A        4022 bp DNA     circular SYN 29-JUL-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4022)
  AUTHORS   Neuroscience
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2022 from SnapGene Viewer 6.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4022
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="M13 fwd
                     common sequencing primer, one of multiple similar variants"
                     /note="color: #a020f0; direction: RIGHT"
     CDS             complement(439..465)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ fragment
                     /note="lacZ-alpha"
                     /note="color: #993366"
                     /translation="ISSLSIPS"
     misc_feature    439..465
                     /label=MCS
                     /note="MCS
                     pBluescript multiple cloning site"
                     /note="color: #a6acb3"
     primer_bind     439..450
                     /label=KS primer
                     /note="KS primer
                     common sequencing primer, one of multiple similar variants"
                     /note="color: #a020f0; direction: RIGHT"
     CDS             complement(471..524)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99b2
                     Cleavage site after base 473"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(549..1271)
                     /codon_start=1
                     /label=cpGFP
                     /note="color: #00ff00"
                     /translation="NVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSVQSILSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPIQVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYN"
     misc_feature    complement(1278..1685)
                     /label=CBD(A135Y)
                     /note="color: #808080; direction: LEFT"
     CDS             complement(1692..1757)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /note="color: #cc99ff
                     Cleavage site after base 1691"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      1757..1766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             complement(1774..1785)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ fragment
                     /note="lacZ-alpha"
                     /note="color: #993366"
                     /translation="LVD"
     misc_feature    1774..1785
                     /label=MCS
                     /note="MCS
                     pBluescript multiple cloning site"
                     /note="color: #a6acb3"
     primer_bind     complement(1774..1785)
                     /label=SK primer
                     /note="SK primer
                     common sequencing primer, one of multiple similar variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(1801..1817)
                     /label=M13 rev
                     /note="M13 rev
                     common sequencing primer, one of multiple similar variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    1825..1841
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator
                     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(1849..1879)
                     /note="lac promoter
                     promoter for the E. coli lac operon"
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    1894..1915
                     /label=E. coli catabolite activator protein binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site
                     CAP binding activates transcription in the presence
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(2203..2791)
                     /direction=LEFT
                     /label=ori
                     /note="ori
                     high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(2962..3822)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR
                     confers resistance to ampicillin, carbenicillin, and 
                     related antibiotics"
                     /note="color: #99ccff"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3823..3927)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tcccaatgtc gacggtatcg ataagcttga tatcgaattc agggccaggg
      481 ttttcctcca cgtcgccgca ggtcagcagg ctgcctctgc cctcagatcc ctcgacaagc
      541 ttactagtgt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      601 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      661 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggatgtagcc ttcgggcatg
      721 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
      781 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
      841 atgaacttca gggtcagctt gccgtaggtg gcatcgccct caccctcgcc ggacacgctg
      901 aacttgtggc cgtttacgtc gccgtccagc tcgacctgga tgggcaccac cccggtgaac
      961 agctcctcgc ccttgctcac catgctccct ccggtaccgc ccttgtacag ctcgtccatg
     1021 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     1081 ttggggtctt tcgaaagaat ggactgcacg ctcaggtagt ggttgtcggg cagcagcacg
     1141 gggccgtcgc cgatgggggt gttctgctgg tagtggtagg cgagctgcac gccgccgtcc
     1201 tcgatgttgt ggcggatctt gaagttcgcc ttgatgccgt tcttctgctt gtcggccttg
     1261 atatagacgt tctcgaggta caggttcccc accttctcag aggtaacctg aagccttctc
     1321 gccatctggg ctgagaggcg catgagtatg tcgggattca cctgtatgag ttgccggaat
     1381 ttcttgtatg agatttcggc cacctcacag gctgtcttag ccctcaccca agcggatctc
     1441 tcctgtcctt cctcgaacag gcccagttct ccgataaagt caccctggtt cagatagctc
     1501 aggatcattt cctttccttc ttcatcctta atcagcacgg cgactgaacc cttcactata
     1561 taatacaggg tttcagcctt ctcgccctga tggatcaggg tgctcttaga tgggtacttg
     1621 tgaatgtgac agtgtgacag aaaccactcc aatgttgggt cggtctgtgg tttgcccagc
     1681 accatggatc ccttgtcatc gtcatccttg taatcgatgt catgatcttt ataatcaccg
     1741 tcatggtctt tgtagtccat ggtggcacta gtagatccac tagttctagt agatctcatg
     1801 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1861 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1921 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1981 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2041 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2101 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2161 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2221 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2281 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2341 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2401 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2461 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2521 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2581 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2641 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2701 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2761 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2821 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2881 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2941 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3001 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3061 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3121 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3181 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3241 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3301 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3361 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3421 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3481 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3541 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3601 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3661 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3721 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3781 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3841 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3901 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3961 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4021 tc
//
