LOCUS       Exported                4516 bp ds-DNA     circular SYN 30-JAN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4516)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Feb 5, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4516
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             174..274
                     /label=HSV TK PolyA
     rep_origin      577..1249
                     /label=pUC origin
                     /note="/ApEinfo_fwdcolor=cyan; /ApEinfo_revcolor=cyan; 
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {0 1 2 0 
                     0 -1} 0} width 5 offset 0"
     enhancer        1366..1669
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1670..1873
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     5'UTR           1901..1943
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     misc_feature    1943..1954
                     /label=clotech kozak
                     /note="Regarding the Kozak,yeah—this is a clotech kozak 
                     (﻿GCAGATACCATG) rather than a consensus.This was originally
                     built this way for historical purposes, but this 
                     slightlyweaker kozak has actually been a benefit in my 
                     hands in that the expressionlevel of the construct is 
                     usable at a more reasonable time window (12-24hours). If 
                     the expression of luminal content gets too high the cells 
                     will dieor get really weird—it depends on how efficiently 
                     your construct is made whenthis will happen (but all 
                     luminal constructs do this eventually…its why stablelumen 
                     cells are really hard to select and usually look weird). I 
                     have oneluminal GFP construct with a consensus kozak and it
                     has to be imaged a littleearlier (the cells are all weird 
                     by 22 hours or so). Unfortunately, the onlyway to really 
                     know this is to test it directly. "
     CDS             1952..2046
                     /label=Prolactin Signal Sequence
     CDS             2054..2428
                     /codon_start=1
                     /product="small protein tag, derived from photoactive 
                     yellow protein, that allows in vivo fluorescent labeling by
                     addition of a cell-permeant fluorogen (Plamont et al., 
                     2016)"
                     /label=Y-FAST
                     /translation="MEHVAFGSEDIENTLANMDDEQLDRLAFGVIQLDGDGNILLYNAA
                     EGDITGRDPKQVIGKNFFKDVAPGTDTPEFYGKFKEGAASGNLNTMFEWTIPTSRGPTK
                     VKVHLKKALSGDRYWVFVKRV"
     CDS             2435..2464
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2620..2690
                     /label=SV40 polyadenylation signal
     rep_origin      complement(2737..3193)
                     /direction=LEFT
                     /label=f1 Origin of Replication
     promoter        3219..3323
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3325..3682
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3717..4511
                     /codon_start=1
                     /transl_table=11
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /db_xref="GI:1377916"
                     /protein_id="AAB02577.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
ORIGIN
        1 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
       61 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
      121 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
      181 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
      241 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
      301 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
      361 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
      421 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
      481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      721 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     1261 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     1321 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     1381 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     1441 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     1501 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     1561 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     1621 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     1681 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     1741 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     1801 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1861 tataagcaga gctggtttag tgaaccgtca gatccgctag cttgttcttt ttgcagaagc
     1921 tcagaataaa cgctcaactt tggcagatac catggacagc aaaggttcgt cgcagaaagg
     1981 gtcccgcctg ctcctgctgc tggtggtgtc aaatctactc ttgtgccagg gtgtggtctc
     2041 ctgcagtcga cttatggagc atgttgcctt tggcagtgag gacatcgaga acactctggc
     2101 caatatggac gacgaacaac tggataggtt ggcctttggc gtaattcagc tcgatggtga
     2161 cgggaatatc ctgctgtaca atgctgctga aggggacatc actggcagag atcccaaaca
     2221 ggtgattggg aagaacttct tcaaggatgt tgcacctgga acggatactc ccgagtttta
     2281 cggcaaattc aaggaaggcg cagcgtcagg gaatctgaac accatgttcg aatggacgat
     2341 accgacaagc aggggaccaa ccaaggtcaa ggtgcacttg aagaaagccc tttccggtga
     2401 cagatattgg gtctttgtga aacgggtggg atccgaacaa aagcttattt ctgaagagga
     2461 cttgaaagat gagctctaag gatccaccgg atctagataa ctgatcataa tcagccatac
     2521 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2581 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2641 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2701 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     2761 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2821 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2881 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2941 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3001 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3061 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3121 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3181 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3241 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3301 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3361 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3421 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3481 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3541 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3601 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3661 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3721 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3781 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3841 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     3901 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3961 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4021 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4081 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4141 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4201 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4261 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4321 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4381 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4441 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4501 agttcttctg agcggg
//
