LOCUS       Exported                4198 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTA-attB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4198)
  AUTHORS   Groth AC, Olivares EC, Thyagarajan B, Calos MP
  TITLE     A phage integrase directs efficient site-specific integration in 
            human cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 May 23. 97(11):5995-6000.
  PUBMED    10801973
REFERENCE   2  (bases 1 to 4198)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4198
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    258..327
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(420..439)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(421..439)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(446..463)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(446..462)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(455..477)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      604..984
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(691..710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     901..922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1352..2146
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1406..1425)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      3195..3783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3684..3703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3937..3954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4071..4092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4107..4137
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4145..4161
                     /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     4150..4172
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4169..4185
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4169..4185
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4181..591
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLVPSSDPLVTAASVLEFGLAVDMPAVTVENPLTLPRVS
                     APADAVARPVLTVTTAPSGFEGEGFPVRRAFAGINYRHLDPFIMMDQMGEVEYAPGEPK
                     GTPWHPHRGFETVTYIVDGTWNSADIHHTGGRSSMHLEGPIRPIVSRITIHWPSFYNVV
                     TGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
ORIGIN
        1 agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga attcggcttg
       61 gctgtcgaca tgcccgccgt gaccgtcgag aacccgctga cgctgccccg cgtatccgca
      121 cccgccgacg ccgtcgcacg tcccgtgctc accgtgacca ccgcgcccag cggtttcgag
      181 ggcgagggct tcccggtgcg ccgcgcgttc gccgggatca actaccgcca cctcgacccg
      241 ttcatcatga tggaccagat gggtgaggtg gagtacgcgc ccggggagcc caagggcacg
      301 ccctggcacc cgcaccgcgg cttcgagacc gtgacctaca tcgtcgacgg tacctggaat
      361 tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg cccaattcgc
      421 cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      481 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
      541 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
      601 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      661 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      721 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      781 ttagagcttt acggcacctc gaccgcaaaa aacttgattt gggtgatggt tcacgtagtg
      841 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      901 gtggactctt gttccaaact ggaacaacac tcaaccctat cgcggtctat tcttttgatt
      961 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaattc
     1021 agggcgcaag ggctgctaaa ggaaccggaa cacgtagaaa gccagtccgc agaaacggtg
     1081 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa
     1141 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg
     1201 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     1261 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc
     1321 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     1381 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     1441 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     1501 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     1561 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     1621 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcgcc ttgctcctgc
     1681 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     1741 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     1801 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     1861 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga tccatggcga
     1921 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca acgactgtgg
     1981 ccggctgggt gtggcggacc gctatcagga catagcgttg gatacccgtg atattgctga
     2041 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     2101 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaattg aaaaaggaag
     2161 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2221 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2281 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2341 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtca tacactatta
     2401 tcccgtattg acgccgggca agagcaactc ggtcgccggg cgcggtattc tcagaatgac
     2461 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2521 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2581 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2641 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagag tgacaccacg
     2701 atgcctgtag caatgccaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2761 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2821 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2881 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2941 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3001 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3301 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     3601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3901 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3961 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     4021 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     4081 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     4141 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgcca
//
