LOCUS       sequence-443108-        6849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Native rat lactase gene (truncated) as an autologous marker (X-Gal, 
            X-Fuc).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6849)
  TITLE     Gene Tracking
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: icLCT
FEATURES             Location/Qualifiers
     source          1..6849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    424..545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(435..454)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     489..508
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    673..744
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     primer_bind     complement(792..809)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(792..808)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(801..823)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1017..1036)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1136..1158
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1196..1214)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1282..1386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1387..1455,1456..2247)
                     /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(1605..1624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2418..3006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2907..2926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3160..3177
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3294..3315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3330..3335,3336..3353,3354..3360)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3368..3384
                     /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     3373..3395
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3392..3408
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3392..3408
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    3450..3521
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     regulatory      4128..4137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgtctc cagaagagga gagcctgaac tcaatgacac tgacaggatc tactacctga
       61 gaagctacat caatgaagcc ctcaaggctg tacaggacaa ggtggacctt agaggctaca
      121 ctgtatggag catcatggac aactttgaat gggccacagg ctttgcagag aggtttggtg
      181 tgcactttgt gaacaggtct gacccatctt tgccaaggat tcccaaggca tcagccaagt
      241 tctatgccac cattgtcaga tgcaatggct ttcctgaccc tgcacaagga cctcatcctt
      301 gtctccagca accagaagat gctgcaccca ctgccagtcc tgtgcagtct gaagtgccct
      361 tcctgggtct gatgctgggc attgcagaag cacagacagc tctgtaaagc ttggatccca
      421 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      481 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      541 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
      601 ttttttaaag caagtaaaac ctctacaaat gtggtaaagg aattcaattc catctctctt
      661 aaggtagcca gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
      721 tcatcaaact ttcttccctc cctaatctat cagatatcta gatgcattcg cgaggtaccg
      781 agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      841 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
      901 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
      961 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     1021 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     1081 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     1141 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     1201 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     1261 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1321 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1381 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1441 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1501 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1561 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1621 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1681 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1741 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1801 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1861 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1921 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1981 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2041 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2101 ggatctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2161 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2221 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2281 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2341 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2401 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2461 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2521 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2581 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2641 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2701 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2761 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2821 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2881 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2941 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3001 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3061 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3121 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3181 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3241 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     3301 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     3361 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     3421 aagcttgcat gcaggcctac atctagatac atcttggacc attagctcca caggtatctt
     3481 cttccctcta gtggtcataa cagcagcttc agctacctct caatcacaac ctaggacatg
     3541 tttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     3601 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     3661 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     3721 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     3781 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     3841 atgaattggt ttgatctgat tataactagt ggagaagagc atgcttgagg gctgagtgcc
     3901 cctcagtggg cagagagcac atggcccaca gtccctgaga agttgggggg aggggtgggc
     3961 aattgaactg gtgcctagag aaggtggggc ttgggtaaac tgggaaagtg atgtggtgta
     4021 ctggctccac ctttttcccc agggtggggg agaaccatat ataagtgcag tagtctctgt
     4081 gaacattcgc tagcagagac ctaagctggc tctcgagaat agctgcagcc accatgggat
     4141 tctcagccaa gaaagtcaag agaaacccac tgcctgtgag agcagacttc acctccagag
     4201 ccagggtcac tgacagcctt ccatctgagg tgccctccaa ggctaagata gtgtgggaga
     4261 agttctccaa gcagcccagg tttgaaagag acttgttcta tcatggcaca ttcagagatg
     4321 acttcctgtg gggtgtatcc tcttcagcct atcagattga aggaggatgg aatgctgatg
     4381 gcaaaggtcc cagcatctgg gataacttca cccacactcc agggaatggt gtgaaagaca
     4441 atgccactgg agatgtagct tgtgacagct atcaccaact ggatgctgac ctaaacatcc
     4501 ttagaaccct gaaggtcaag tcctatagat tctccatctc ctggtccaga atcttcccaa
     4561 caggaagaaa cagtaccatc aacaaacaag gtgttgatta ttacaacaga ctgattgaca
     4621 gcttggtgga caacaacatc ttccctatgg tgacactgtt tcactgggac ctgcctcagg
     4681 ccctccagga tattggaggt tgggagaacc cttctttgat tgagttgttt gacagctatg
     4741 cagacttctg ctttaagacc tttggtgaca gagtcaagtt ctggatgacc tttaatgagc
     4801 cctggtgtca tgtggtgtta ggctacagtt cagggatatt tcctccaagt gttcaggagc
     4861 caggctggct accttataag gttagccata ttgtcattaa ggcacatgct agggtctacc
     4921 atacttatga tgagaagtac aggtctgagc agaaaggtgt catctctctc agcctcaaca
     4981 ctcactgggc agaacccaag gacccagggc tccaaaggga tgtggaagct gctgacagaa
     5041 tgctgcagtt tactatgggc tggtttgccc accctatctt caagaatggt gactatccag
     5101 atgtgatgaa atggactgtg ggtaacagga gtgagctgca gcacctggcc agctccagac
     5161 tgcctacctt cacagaggaa gagaagaatt atgtcagagg cactgctgat gtcttctgca
     5221 tcaacaccta cacttctgtg tttgtgcagc acagcactcc caggctgaac ccacccagct
     5281 atgatgatga catggagttg aagctaattg aaatgaactc ttctactggt gtgatgcacc
     5341 cagatgtgcc ctggggtact aggaggctgc tgaactggat caaggaggag tatggcaaca
     5401 ttcctattta tatcactgag aatggacagg gactggagaa tcccacactg gatgacacag
     5461 aaaggatctt ctaccacaag acctatatca atgaggctct gaaagcctac aagctggatg
     5521 gtgtggacct tagagggtac tctgcctgga ctctgatgga tgactttgag tggctgcttg
     5581 gctacaccat gagatttgga ttgtatcatg ttgactttaa tcatgtgagc agacctagaa
     5641 cagcaagagc ctcagccaga tactatgcag aggtcattgc caacaatggt atgcctctgg
     5701 ccagggaaga tgagttcctg tatggagagt ttcccaaggg cttcatctgg agtgctgctt
     5761 ctgcttcata tcaggttgaa ggtgcttgga gagcagatgg caaaggactc agtatctggg
     5821 acacattctc ccacacacca ctgaggattg ggaatgatga caatggagat gtggcatgtg
     5881 acagttacca caagattgct gaggatgtgg tggcccttca gaacctgggt gtgtcccact
     5941 atagattctc cattgcctgg tccagaatcc tcccagatgg aaccaccaag ttcatcaatg
     6001 aagctggcct cagttactat gttagattca ttgatgccct gctggctgct ggcatcacac
     6061 cacaggtgac tatttatcac tgggacctgc cacaggcact gcaagatgtg ggtggctggg
     6121 agaatgagac tattgtgcag agatttaagg agtatgctga tgttctcttc cagaggctgg
     6181 gtgacagagt caagttctgg atcactctga atgagccctt tgtcattgca gcccaaggct
     6241 atggcactgg tgtgtctgct ccaggaatct ccttcagacc tggcacagct ccttacattg
     6301 ctggacacaa cctaataaag gcccatgctg aagcctggca tctctacaat gatgtataca
     6361 gagccagaca gggaggcacc atctccatca ctatcagcag tgactgggct gaacccagag
     6421 atcccaccaa ccagggagat gtggaggcag ccaggagata tgtacagttt atgggtggct
     6481 ggtttgctca ccctatcttc aagaatggag attaccctga ggtgatgaag acaagaatca
     6541 gagacagaag cctggctgct ggcctcaaca agtccaggct ccctgagttt actgagagtg
     6601 agaagagtag gatcaagggc acctttgact tctttggttt caaccactac accactgtcc
     6661 tagcttacaa ccttgactac cctgctgcct tctcctcctt tgatgcagac aggggtgttg
     6721 cttccattgc agacagctct tggccagtct ctggctcctt ctggctgaag gtgactccat
     6781 ttggcttcag aaggatcttg aactggctga aggaggagta caataatcct cctatttatg
     6841 tcactgaga
//
